US20080299618A1 - Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors - Google Patents
Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors Download PDFInfo
- Publication number
- US20080299618A1 US20080299618A1 US12/127,237 US12723708A US2008299618A1 US 20080299618 A1 US20080299618 A1 US 20080299618A1 US 12723708 A US12723708 A US 12723708A US 2008299618 A1 US2008299618 A1 US 2008299618A1
- Authority
- US
- United States
- Prior art keywords
- sequence
- primers
- variable domain
- dna
- primer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/655—Somatostatins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/461—Igs containing Ig-regions, -domains or -residues form different species
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
Definitions
- the present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- Ig immunoglobulin
- FIG. 1 shows a schematic representation of the unrearranged and rearranged heavy and light chain variable genes and the location of the primers.
- FIG. 2 shows a schematic representation of the M13-VHPCR1 vector and a cloning scheme for amplified heavy chain variable domains.
- FIG. 3 shows the sequence of the Ig variable region derived sequences in M13-VHPCR1.
- FIG. 4 shows a schematic representation of the M13-VKPCR1 vector and a cloning scheme for light chain variable domains.
- FIG. 5 shows the sequence of the Ig variable region derived sequences in M13-VKPCR1.
- FIG. 6 shows the nucleotide sequences of the heavy and light chain variable domain encoding sequences of MAb MBr1.
- FIG. 7 shows a schematic representation of the pSV-gpt vector (also known as ⁇ -Lys 30) which contains a variable region cloned as a HindIII-BamHI fragment, which is excised on introducing the new variable region.
- the gene for human IgG1 has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique.
- FIG. 8 shows a schematic representation of the pSV-hygro vector (also known as ⁇ -Lys 17). It is derived from pSV gpt vector with the gene encoding mycophenolic acid replaced by a gene coding for hygromycin resistance. The construct contains a variable gene cloned as a HindIII-BamHI fragment which is excised on introducing the new variable region. The gene for human C ⁇ has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique.
- FIG. 9 shows the assembly of the mouse: human MBr1 chimeric antibody.
- FIGS. 10 a - 10 b shows encoded amino acid sequences of 48 mouse rearranged VH genes.
- FIG. 11 shows encoded amino acid sequences of human rearranged VH genes.
- FIG. 12 shows encoded amino acid sequences of unrearranged human VH genes.
- FIG. 13 shows the sequence of part of the plasmid pSW1: essentially the sequence of a pectate lyase leader linked to VHLYS in pSW1 and cloned as an SphI-EcoRI fragment into pUC19 and the translation of the open reading frame encoding the pectate lyase leader-VHLYS polypeptide being shown.
- FIGS. 14 a - 14 b shows the sequence of part of the plasmid pSW2: essentially the sequence of a pectate lyase leader linked to VHLYS and to VKLYS, and cloned as an SphI-EcoRI-EcoRI fragment into pUC19 and the translation of open reading frames encoding the pectate lyase leader-VHLYS and pectate lyase leader-VKLYS polypeptides being shown.
- FIG. 15 shows the sequence of part of the plasmid pSW1HPOLYMYC which is based on pSW1 and in which a polylinker sequence has replaced the variable domain of VHLYS, and acts as a cloning site for amplified VH genes, and a peptide tag is introduced at the C-terminal end.
- FIG. 16 shows the encoded amino acid sequences of two VH domains derived from mouse spleen and having lysozyme binding activity, and compared with the VH domain of the D1,3 antibody.
- the arrows mark the points of difference between the two VH domains.
- FIG. 17 shows the encoded amino acid sequence of a VH domain derived from human peripheral blood lymphocytes and having lysozyme binding activity.
- FIG. 18 shows a scheme for generating and cloning mutants of the VHLYS gene, which is compared with the scheme for cloning natural repertoires of VH genes.
- FIG. 19 shows the sequence of part of the vector pSW2HPOLY.
- FIG. 20 shows the sequence of part of the vector pSW3 which encodes the two linked VHLYS domains.
- FIGS. 21 a - 21 c shows the sequence of the VHLYS domain and pelB leader sequence fused to the alkaline phosphatase gene.
- FIG. 22 shows the sequence of the vector pSW1VHLYS-VKPOLYMYC for expression of a repertoire of V ⁇ light chain variable domains in association with the VHLYS domain.
- FIG. 23 shows the sequence of VH domain which is secreted at high levels from E. coli . The differences with VHLYS domain are marked.
- the present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- Ig immunoglobulin
- the Ig superfamily includes not only the Igs themselves but also such molecules as receptors on lymphoid cells such as T lymphocytes.
- Immunoglobulins comprise at least one heavy and one light chain covalently bonded together. Each chain is divided into a number of domains. At the N-terminal end of each chain is a variable domain. The variable domains on the heavy and light chains fit together to form a binding site designed to receive a particular target molecule. In the case of Igs, the target molecules are antigens.
- T-cell receptors have two chains of equal size, the ⁇ and ⁇ chains, each consisting of two domains.
- variable domains on the ⁇ and ⁇ chains are believed to fit together to form a binding site for target molecules, in this case peptides presented by a histocompatibility antigen.
- the variable domains are so called because their amino acid sequences vary particularly from one molecule to another. This variation in sequence enables the molecules to recognize an extremely wide variety of target molecules.
- each variable domain comprises a number of areas of relatively conserved sequence and three areas of hypervariable sequence.
- the three hypervariable areas are generally known as complementarity determining regions (CDRs).
- Boss The Boss application also relates to the cloning and expression of chimeric antibodies.
- Chimeric antibodies are Ig-type molecules in which the variable domains from one Ig are fused to constant domains from another Ig.
- the variable domains are derived from an Ig from one species (often a mouse Ig) and the constant domains are derived from an Ig from a different species (often a human Ig).
- EP-A-0 125 023 (Genentech) relates to much the same subject as the Boss application, but also relates to the production by recombinant DNA technology of other variations of Ig-type molecules.
- EP-A-0 194 276 discloses not only chimeric antibodies of the type disclosed in the Boss application but also chimeric antibodies in which some or all of the constant domains have been replaced by non-Ig derived protein sequences.
- the heavy chain CH2 and CH3 domains may be replaced by protein sequences derived from an enzyme or a protein toxin.
- EP-A-0 239 400 discloses a different approach to the production of Ig molecules.
- this approach only the CDRs from a first type of Ig are grafted onto a second type of Ig in place of its normal CDRs.
- the Ig molecule thus produced is predominantly of the second type, since the CDRs form a relatively small part of the whole Ig.
- the CDRs are the parts which define the specificity of the Ig, the Ig molecule thus produced has its specificity derived from the first Ig.
- modified antibodies chimeric antibodies, CDR-grafted Igs, the altered antibodies described by Genentech, and fragments of such Igs such as F(ab′) 2 and Fv fragments are referred to herein as modified antibodies.
- MAbs monoclonal antibodies
- MAbs directed against cancer antigens have been produced. It is envisaged that these MAbs could be covalently attached or fused to toxins to provide “magic bullets” for use in cancer therapy. MAbs directed against normal tissue or cell surface antigens have also been produced. Labels can be attached to these so that they can be used for in vivo imaging.
- MAbs in therapy or in vivo diagnosis
- the vast majority of MAbs which are produced are of rodent, in particular mouse, origin. It is very difficult to produce human MAbs. Since most MAbs are derived from non-human species, they are antigenic in humans. Thus, administration of these MAbs to humans generally results in an anti-Ig response being mounted by the human. Such a response can interfere with therapy or diagnosis, for instance by destroying or clearing the antibody quickly, or can cause allergic reactions or immune complex hypersensitivity which has adverse effects on the patient.
- modified Igs have been proposed to ensure that the Ig administered to a patient is as “human” as possible, but still retains the appropriate specificity. It is therefore expected that modified Igs will be as effective as the MAb from which the specificity is derived but at the same time not very antigenic. Thus, it should be possible to use the modified Ig a reasonable number of times in a treatment or diagnosis regime.
- heavy chain variable domains are encoded by a “rearranged” gene which is built from three gene segments: an “unrearranged” VH gene (encoding the N-terminal three framework regions, first two complete CDRs and the first part of the third CDR), a diversity (DH)-segment (DH) (encoding the central portion of the third CDR) and a joining segment (JH) (encoding the last part of the third CDR and the fourth framework region).
- VH gene encoding the N-terminal three framework regions, first two complete CDRs and the first part of the third CDR
- DH diversity-segment
- JH joining segment
- light chain variable domains are encoded by an “unrearranged” VL gene and a JL gene.
- VL gene There are two types of light chains, kappa ( ⁇ ) or lambda ( ⁇ ), which are built respectively from unrearranged V ⁇ genes and J ⁇ segments, and from unrearranged V ⁇ genes and J ⁇ segments.
- Ig heavy chain variable domains can bind to antigen in a 1:1 ratio and with binding constants of equivalent magnitude to those of complete antibody molecules. In view of what was known up until now and in view of the assumptions made by those skilled in the art, this is highly surprising.
- a single domain ligand consisting of at least part of the variable domain of one chain of a molecule from the Ig superfamily.
- the ligand consists of the variable domain of an Ig light, or, most preferably, heavy chain.
- the ligand may be produced by any known technique, for instance by controlled cleavage of Ig superfamily molecules or by peptide synthesis. However, preferably the ligand is produced by recombinant DNA technology. For instance, the gene encoding the rearranged gene for a heavy chain variable domain may be produced, for instance by cloning or gene synthesis, and placed into a suitable expression vector. The expression vector is then used to transform a compatible host cell which is then cultured to allow the ligand to be expressed and, preferably, secreted.
- the gene for the ligand can be mutated to improve the properties of the expressed domain, for example to increase the yields of expression or the solubility of the ligand, to enable the ligand to bind better, or to introduce a second site for covalent attachment (by introducing chemically reactive residues such as cysteine and histidine) or non-covalent binding of other molecules.
- a second site for binding to serum components to prolong the residence time of the domains in the serum; or for binding to molecules with effector functions, such as components of complement, or receptors on the surfaces of cells.
- hydrophobic residues which would normally be at the interface of the heavy chain variable domain with the light chain variable domain could be mutated to more hydrophilic residues to improve solubility; residues in the CDR loops could be mutated to improve antigen binding; residues on the other loops or parts of the ⁇ -sheet could be mutated to introduce new binding activities. Mutations could include single point mutations, multiple point mutations or more extensive changes and could be introduced by any of a variety of recombinant DNA methods, for example gene synthesis, site directed mutagenesis or the polymerase chain reaction.
- the ligands of the present invention have equivalent binding affinity to that of complete Ig molecules
- the ligands can be used in many of the ways as are Ig molecules or fragments.
- Ig molecules have been used in therapy (such as in treating cancer, bacterial and viral diseases), in diagnosis (such as pregnancy testing), in vaccination (such as in producing anti-idiotypic antibodies which mimic antigens), in modulation of activities of hormones or growth factors, in detection, in biosensors and in catalysis.
- the small size of the ligands of the present invention may confer some advantages over complete antibodies, for example, in neutralizing the activity of low molecular weight drugs (such as dioxin) and allowing their filtration from the kidneys with drug attached; in penetrating tissues and tumors; in neutralizing viruses by binding to small conserved regions on the surfaces of viruses such as the “canyon” sites of viruses [16]; in high resolution epitope mapping of proteins; and in vaccination by ligands which mimic antigens.
- low molecular weight drugs such as dioxin
- the present invention also provides receptors comprising a ligand according to the first aspect of the invention linked to one or more of an effector molecule, a label, a surface, or one or more other ligands having the same or different specificity.
- a receptor comprising a ligand linked to an effector molecule may be of use in therapy.
- the effector molecule may be a toxin, such as ricin or pseudomonas exotoxin, an enzyme which is able to activate a prodrug, a binding partner or a radio-isotope.
- the radio-isotope may be directly linked to the ligand or may be attached thereto by a chelating structure which is directly linked to the ligand.
- Such ligands with attached isotopes are much smaller than those based on Fv fragments, and could penetrate tissues and access tumors more readily.
- a receptor comprising a ligand linked to a label may be of use in diagnosis.
- the label may be a heavy metal atom or a radio-isotope, in which case the receptor can be used for in vivo imaging using X-ray or other scanning apparatus.
- the metal atom or radio-isotope may be attached to the ligand either directly or via a chelating structure directly linked to the ligand.
- the label may be a heavy metal atom, a radio-isotope, an enzyme, a fluorescent or colored molecule or a protein or peptide tag which can be detected by an antibody, an antibody fragment or another protein.
- Such receptors would be used in any of the known diagnostic tests, such as ELISA or fluorescence-linked assays.
- a receptor comprising a ligand linked to a surface could be used for purification of other molecules by affinity chromatography.
- Linking of ligands to cells for example to the outer membrane proteins of E. coli or to hydrophobic tails which localize the ligands in the cell membranes, could allow a simple diagnostic test in which the bacteria or cells would agglutinate in the presence of molecules bearing multiple sites for binding the ligand(s).
- Receptors comprising at least two ligands can be used, for instance, in diagnostic tests.
- the first ligand will bind to a test antigen and the second ligand will bind to a reporter molecule, such as an enzyme, a fluorescent dye, a colored dye, a radio-isotope or a colored-, fluorescently- or radio-labelled protein.
- such receptors may be useful in increasing the binding to an antigen.
- the first ligand will bind to a first epitope of the antigen and the second ligand will bind to a second epitope.
- Such receptors may also be used for increasing the affinity and specificity of binding to different antigens in close proximity on the surface of cells.
- the first ligand will bind to the first antigen and the second epitope to the second antigen: strong binding will depend on the co-expression of the epitopes on the surface of the cell. This may be useful in therapy of tumors, which can have elevated expression of several surface markers. Further ligands could be added to further improve binding or specificity.
- the use of strings of ligands with the same or multiple specificities, creates a larger molecule which is less readily filtered from the circulation by the kidney.
- the use of strings of ligands may prove more effective than single ligands, due to repetition of the immunizing epitopes.
- such receptors with multiple ligands could include effector molecules or labels so that they can be used in therapy or diagnosis as described above.
- the ligand may be linked to the other part of the receptor by any suitable means, for instance by covalent or non-covalent chemical linkages.
- the receptor comprises a ligand and another protein molecule, it is preferred that they are produced by recombinant DNA technology as a fusion product. If necessary, a linker peptide sequence can be placed between the ligand and the other protein molecule to provide flexibility.
- the ligand is to be used for in vivo diagnosis or therapy in humans, it is humanized, for instance by CDR replacement as described in EP-A-0 239 400.
- a further problem with the production of ligands, and also receptors according to the invention and modified Igs, by recombinant DNA technology is the cloning of the variable domain encoding sequences from the hybridoma which produces the MAb from which the specificity is to be derived.
- This can be a relatively long method involving the production of a suitable probe, construction of a clone library from cDNA or genomic DNA, extensive probing of the clone library, and manipulation of any isolated clones to enable the cloning into a suitable expression vector. Due to the inherent variability of the DNA sequences encoding Ig variable domains, it has not previously been possible to avoid such time consuming work. It is therefore a further aim of the present invention to provide a method which enables substantially any sequence encoding an Ig superfamily molecule variable domain (ligand) to be cloned in a reasonable period of time.
- a method of cloning a sequence (the target sequence) which encodes at least part of the variable domain of an Ig superfamily molecule which method comprises:
- a forward and a back oligonucleotide primer annealing to the sample a forward and a back oligonucleotide primer, the forward primer being specific for a sequence at or adjacent the 3′ end of the sense strand of the target sequence, the back primer being specific for a sequence at or adjacent the 3′ end of the antisense strand of the target sequence, under conditions which allow the primers to hybridize to the nucleic acid at or adjacent the target sequence;
- the method of the present invention further includes the step (f) of repeating steps (c) to (e) on the denatured mixture a plurality of times.
- the method of the present invention is used to clone complete variable domains from Ig molecules, most preferably from Ig heavy chains.
- the method will produce a DNA sequence encoding a ligand according to the present invention.
- step (c) recited above the forward primer becomes annealed to the sense strand of the target sequence at or adjacent the 3′ end of the strand.
- the back primer becomes annealed to the antisense strand of the target sequence at or adjacent the 3′ end of the strand.
- the forward primer anneals at or adjacent the region of the ds nucleic acid which encodes the C-terminal end of the variable region or domain.
- the back primer anneals at or adjacent the region of the ds nucleic acid which encodes the N-terminal end of the variable domain.
- step (d) nucleotides are added onto the 3′ end of the forward and back primers in accordance with the sequence of the strand to which they are annealed. Primer extension will continue in this manner until stopped by the beginning of the denaturing step (e). It must therefore be ensured that step (d) is carried out for a long enough time to ensure that the primers are extended so that the extended strands totally overlap one another.
- step (e) the extended primers are separated from the ds nucleic acid.
- the ds nucleic acid can then serve again as a substrate to which further primers can anneal.
- the extended primers themselves have the necessary complementary sequences to enable the primers to anneal thereto.
- step (f) the amount of extended primers will increase exponentially so that at the end of the cycles there will be a large quantity of cDNA having sequences complementary to the sense and antisense strands of the target sequence.
- the method of the present invention will result in the accumulation of a large quantity of cDNA which can form ds cDNA encoding at least part of the variable domain.
- the forward and back primers may be provided as isolated oligonucleotides, in which case only two oligonucleotides will be used. However, alternatively the forward and back primers may each be supplied as a mixture of closely related oligonucleotides. For instance, it may be found that at a particular point in the sequence to which the primer is to anneal, there is the possibility of nucleotide variation. In this case a primer may be used for each possible nucleotide variation. Furthermore it may be possible to use two or more sets of “nested” primers in the method to enhance the specific cloning of variable region genes.
- RNA may be isolated in known manner from a cell or cell line which is known to produce Igs.
- mRNA may be separated from other RNA by oligo-dT chromatography.
- a complementary strand of cDNA may then be synthesized on the mRNA template, using reverse transcriptase and a suitable primer, to yield an RNA/DNA heteroduplex.
- a second strand of DNA can be made in one of several ways, for example, by priming with RNA fragments of the mRNA strand (made by incubating RNA/DNA heteroduplex with RNase H) and using DNA polymerase, or by priming with a synthetic oligodeoxynucleotide primer which anneals to the 3′ end of the first strand and using DNA polymerase. It has been found that the method of the present invention can be carried out using ds cDNA prepared in this way.
- a forward primer which anneals to a sequence in the CH1 domain (for a heavy chain variable domain) or the C ⁇ or C ⁇ domain (for a light chain variable domain). These will be located in close enough proximity to the target sequence to allow the sequence to be cloned.
- the back primer may be one which anneals to a sequence at the N-terminal end of the VH1, V ⁇ or V ⁇ domain.
- the back primer may consist of a plurality of primers having a variety of sequences designed to be complementary to the various families of VH1, V ⁇ or V ⁇ sequences known.
- the back primer may be a single primer having a consensus sequence derived from all the families of variable region genes.
- the method of the present invention can be carried out using genomic DNA. If genomic DNA is used, there is a very large amount of DNA present, including actual coding sequences, introns and untranslated sequences between genes. Thus, there is considerable scope for non-specific annealing under the conditions used. However, it has surprisingly been found that there is very little non-specific annealing. It is therefore unexpected that it has proved possible to clone the genes of Ig-variable domains from genomic DNA.
- genomic DNA may prove advantageous compared with use of mRNA, as the mRNA is readily degraded, and especially difficult to prepare from clinical samples of human tissue.
- the ds nucleic acid used in step (a) is genomic DNA.
- genomic DNA As the ds nucleic acid source, it will not be possible to use as the forward primer an oligonucleotide having a sequence complementary to a sequence in a constant domain. This is because, in genomic DNA, the constant domain genes are generally separated from the variable domain genes by a considerable number of base pairs. Thus, the site of annealing would be too remote from the sequence to be cloned.
- the method of the present invention can be used to clone both rearranged and unrearranged variable domain sequences from genomic DNA. It is known that in germ line genomic DNA the three genes, encoding the VH, DH and JH respectively, are separated from one another by considerable numbers of base pairs. On maturation of the immune response, these genes are rearranged so that the VH, DH and JH genes are fused together to provide the gene encoding the whole variable domain (see FIG. 1 ). By using a forward primer specific for a sequence at or adjacent the 3′ end of the sense strand of the genomic “unrearranged” VH gene, it is possible to clone the “unrearranged” VH gene alone, without also cloning the DH and JH genes. This can be of use in that it will then be possible to fuse the VH gene onto pre-cloned or synthetic DH and DH genes. In this way, rearrangement of the variable domain genes can be carried out in vitro.
- the oligonucleotide primers used in step (c) may be specifically designed for use with a particular target sequence. In this case, it will be necessary to sequence at least the 5′ and 3′ ends of the target sequence so that the appropriate oligonucleotides can be synthesized. However, the present inventors have discovered that it is not necessary to use such specifically designed primers. Instead, it is possible to use a species specific general primer or a mixture of such primers for annealing to each end of the target sequence. This is not particularly surprising as regards the 3′ end of the target sequence. It is known that this end of the variable domain encoding sequence leads into a segment encoding JH which is known to be relatively conserved. However, it was surprisingly discovered that, within a single species, the sequence at the 5′ end of the target sequence is sufficiently well conserved to enable a species specific general primer or a mixture thereof to be designed for the 5′ end of the target sequence.
- the two primers which are used are species specific general primers, whether used as single primers or as mixtures of primers. This greatly facilitates the cloning of any undetermined target sequence since it will avoid the need to carry out any sequencing on the target sequence in order to produce target sequence-specific primers.
- the method of this aspect of the invention provides a general method for cloning variable region or domain encoding sequences of a particular species.
- variable domain gene Once the variable domain gene has been cloned using the method described above, it may be directly inserted into an expression vector, for instance using the PCR reaction to paste the gene into a vector.
- each primer includes a sequence including a restriction enzyme recognition site.
- the sequence recognized by the restriction enzyme need not be in the part of the primer which anneals to the ds nucleic acid, but may be provided as an extension which does not anneal.
- the use of primers with restriction sites has the advantage that the DNA can be cut with at least one restriction enzyme which leaves 3′ or 5′ overhanging nucleotides. Such DNA is more readily cloned into the corresponding sites on the vectors than blunt end fragments taken directly from the method. The ds cDNA produced at the end of the cycles will thus be readily insertable into a cloning vector by use of the appropriate restriction enzymes.
- restriction sites is such that the ds cDNA is cloned directly into an expression vector, such that the ligand encoded by the gene is expressed.
- the restriction site is preferably located in the sequence which is annealed to the ds nucleic acid.
- the primers may not have a sequence exactly complementary to the target sequence to which it is to be annealed, for instance because of nucleotide variations or because of the introduction of a restriction enzyme recognition site, it may be necessary to adjust the conditions in the annealing mixture to enable the primers to anneal to the ds nucleic acid. This is well within the competence of the person skilled in the art and needs no further explanation.
- any DNA polymerase may be used.
- Such polymerases are known in the art and are available commercially. The conditions to be used with each polymerase are well known and require no further explanation here.
- the polymerase reaction will need to be carried out in the presence of the four nucleoside triphosphates. These and the polymerase enzyme may already be present in the sample or may be provided afresh for each cycle.
- the denaturing step (e) may be carried out, for instance, by heating the sample, by use of chaotropic agents, such as urea or guanidine, or by the use of changes in ionic strength or pH.
- chaotropic agents such as urea or guanidine
- denaturing is carried out by heating since this is readily reversible.
- thermostable DNA polymerase such as Taq polymerase, since this will not need replenishing at each cycle.
- a suitable cycle of heating comprises denaturation at about 95° C. for about 1 minute, annealing at from 30° C. to 65° C. for about 1 minute and primer extension at about 75° C. for about 2 minutes.
- the mixture after the final cycle is preferably held at about 60° C. for about 5 minutes.
- the product ds cDNA may be separated from the mixture for instance by gel electrophoresis using agarose gels.
- the ds cDNA may be used in unpurified form and inserted directly into a suitable cloning or expression vector by conventional methods. This will be particularly easy to accomplish if the primers include restriction enzyme recognition sequences.
- the method of the present invention may be used to make variations in the sequences encoding the variable domains.
- this may be achieved by using a mixture of related oligonucleotide primers as at least one of the primers.
- the primers are particularly variable in the middle of the primer and relatively conserved at the 5′ and 3′ ends.
- the ends of the primers are complementary to the framework regions of the variable domain, and the variable region in the middle of the primer covers all or part of a CDR.
- a forward primer is used in the area which forms the third CDR. If the method is carried out using such a mixture of oligonucleotides, the product will be a mixture of variable domain encoding sequences.
- variations in the sequence may be introduced by incorporating some mutagenic nucleotide triphosphates in step (d), such that point mutations are scattered throughout the target region.
- point mutations are introduced by performing a large number of cycles of amplification, as errors due to the natural error rate of the DNA polymerase are amplified, particularly when using high concentrations of nucleoside triphosphates.
- the method of this aspect of the present invention has the advantage that it greatly facilitates the cloning of variable domain encoding sequences directly from mRNA or genomic DNA. This in turn will facilitate the production of modified Ig-type molecules by any of the prior art methods referred to above. Further, target genes can be cloned from tissue samples containing antibody producing cells, and the genes can be sequenced. By doing this, it will be possible to look directly at the immune repertoire of a patient. This “fingerprinting” of a patient's immune repertoire could be of use in diagnosis, for instance of auto-immune diseases.
- a single set of primers is used in several cycles of copying via the polymerase chain reaction.
- steps (a) to (d) as above which further includes the steps of:
- the second method may further include the steps of:
- fragments are separated from the vector and from other fragments of the incorrect size by gel electrophoresis.
- steps (a) to (d) then (g) to (h) can be followed once, but preferably the entire cycle (c) to (d) and (g) to (h) is repeated at least once.
- a priming step in which the genes are specifically copied, is followed by a cloning step, in which the amount of genes is increased.
- the ds cDNA is derived from mRNA.
- the mRNA is preferably be isolated from lymphocytes which have been stimulated to enhance production of mRNA.
- the set of primers are preferably different from the previous step (c), so as to enhance the specificity of copying.
- the sets of primers form a nested set.
- the first set of primers may be located within the signal sequence and constant region, as described by Larrick et al., [18], and the second set of primers entirely within the variable region, as described by Orlandi et al., [19].
- the primers of step (c) include restriction sites to facilitate subsequent cloning.
- the set of primers used in step (c) should preferably include restriction sites for introduction into expression vectors.
- step (g) possible mismatches between the primers and the template strands are corrected by “nick translation”.
- step (h) the ds cDNA is preferably cleaved with restriction enzymes at sites introduced into the primers to facilitate the cloning.
- the product ds cDNA is cloned directly into an expression vector.
- the host may be prokaryotic or eukaryotic, but is preferably bacterial.
- restriction sites in the primers and in the vector, and other features of the vector will allow the expression of complete ligands, while preserving all those features of the amino acid sequence which are typical of the (methoded) ligands.
- the primers would be chosen to allow the cloning of target sequences including at least all the three CDR sequences.
- the cloning vector would then encode a signal sequence (for secretion of the ligand), and sequences encoding the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the last (fourth) framework region.
- the primers would be chosen to allow the cloning of target sequences including at least the first two CDRs.
- the cloning vector could then encode signal sequence, the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the third framework region, the third CDR and fourth framework region.
- Primers and cloning vectors may likewise be devised for expression of single CDRs, particularly the third CDR, as parts of complete ligands.
- the advantage of cloning repertoires of single CDRs would permit the design of a “universal” set of framework regions, incorporating desirable properties such as solubility.
- Single ligands could be expressed alone or in combination with a complementary variable domain.
- a heavy chain variable domain can be expressed either as an individual domain or, if it is expressed with a complementary light chain variable domain, as an antigen binding site.
- the two partners would be expressed in the same cell, or secreted from the same cell, and the proteins allowed to associate non-covalently to form an Fv fragment.
- the two genes encoding the complementary partners can be placed in tandem and expressed from a single vector, the vector including two sets of restriction sites.
- the genes are introduced sequentially: for example the heavy chain variable domain can be cloned first and then the light chain variable domain.
- the two genes are introduced into the vector in a single step, for example by using the polymerase chain reaction to paste together each gene with any necessary intervening sequence, as essentially described by Yon and Fried [29].
- the two partners could be also expressed as a linked protein to produce a single chain Fv fragment, using similar vectors to those described above.
- the two genes may be placed in two different vectors, for example in which one vector is a phage vector and the other is a plasmid vector.
- the cloned ds cDNA may be inserted into an expression vector already containing sequences encoding one or more constant domains to allow the vector to express Ig-type chains.
- the expression of Fab fragments would have the advantage over Fv fragments that the heavy and light chains would tend to associate through the constant domains in addition to the variable domains.
- the final expression product may be any of the modified Ig-type molecules referred to above.
- the cloned sequence may also be inserted into an expression vector so that it can be expressed as a fusion protein.
- the variable domain encoding sequence may be linked directly or via a linker sequence to a DNA sequence encoding any protein effector molecule, such as a toxin, enzyme, label or another ligand.
- the variable domain sequences may also be linked to proteins on the outer side of bacteria or phage.
- the method of this aspect of the invention may be used to produce receptors according to the invention.
- the cloning of ds cDNA directly for expression permits the rapid construction of expression libraries which can be screened for binding activities.
- the ds cDNA may comprise variable genes isolated as complete rearranged genes from the animal, or variable genes built from several different sources, for example a repertoire of unrearranged VH genes combined with a synthetic repertoire of DH and JH genes.
- repertoires of genes encoding Ig heavy chain variable domains are prepared from lymphocytes of animals immunized with an antigen.
- the screening method may take a range of formats well known in the art.
- Ig heavy chain variable domains secreted from bacteria may be screened by binding to antigen on a solid phase, and detecting the captured domains by antibodies.
- the domains may be screened by growing the bacteria in liquid culture and binding to antigen coated on the surface of ELISA plates.
- bacterial colonies or phage plaques which secrete ligands (or modified ligands, or ligand fusions with proteins) are screened for antigen binding on membranes.
- Either the ligands are bound directly to the membranes (and for example detected with labelled antigen), or captured on antigen coated membranes (and detected with reagents specific for ligands).
- the use of membranes offers great convenience in screening many clones, and such techniques are well known in the art.
- the screening method may also be greatly facilitated by making protein fusions with the ligands, for example by introducing a peptide tag which is recognized by an antibody at the N-terminal or C-terminal end of the ligand, or joining the ligand to an enzyme which catalyses the conversion of a colorless substrate to a colored product.
- the binding of antigen may be detected simply by adding substrate.
- joining of the ligand and a domain of a transcriptional activator such as the GAL4 protein of yeast, and joining of antigen to the other domain of the GAL4 protein, could form the basis for screening binding activities, as described by Fields and Song [21].
- the preparation of proteins, or even cells with multiple copies of the ligands may improve the avidity of the ligand for immobilized antigen, and hence the sensitivity of the screening method.
- the ligand may be joined to a protein subunit of a multimeric protein, to a phage coat protein or to an outer membrane protein of E. coli such as ompA or lamB.
- Such fusions to phage or bacterial proteins also offers possibilities of selecting bacteria displaying ligands with antigen binding activities.
- bacteria may be precipitated with antigen bound to a solid support, or may be subjected to affinity chromatography, or may be bound to larger cells or particles which have been coated with antigen and sorted using a fluorescence activated cell sorter (FACS).
- FACS fluorescence activated cell sorter
- the proteins or peptides fused to the ligands are preferably encoded by the vector, such that cloning of the ds cDNA repertoire creates the fusion product.
- the associated Ig heavy and light chain variable domains For example, repertoires of heavy and light chain variable genes may be cloned such that two domains are expressed together. Only some of the pairs of domains may associate, and only some of these associated pairs may bind to antigen.
- the repertoires of heavy and light chain variable domains could be cloned such that each domain is paired at random. This approach may be most suitable for isolation of associated domains in which the presence of both partners is required to form a cleft. Alternatively, to allow the binding of hapten.
- a small repertoire of light chain variable domains for example including representative members of each family of domains, may be combined with a large repertoire of heavy chain variable domains.
- a repertoire of heavy chain variable domains is screened first for antigen binding in the absence of the light chain partner, and then only those heavy chain variable domains binding to antigen are combined with the repertoire of light chain variable domains. Binding of associated heavy and light chain variable domains may be distinguished readily from binding of single domains, for example by fusing each domain to a different C-terminal peptide tag which are specifically recognized by different monoclonal antibodies.
- the hierarchical approach of first cloning heavy chain variable domains with binding activities, then cloning matching light chain variable domains may be particularly appropriate for the construction of catalytic antibodies, as the heavy chain may be screened first for substrate binding.
- a light chain variable domain would then be identified which is capable of association with the heavy chain, and “catalytic” residues such as cysteine or histidine (or prosthetic groups) would be introduced into the CDRs to stabilize the transition state or attack the substrate, as described by Baldwin and Schultz [22].
- Fab fragments are more likely to be associated than the Fv fragments, as the heavy chain variable domain is attached to a single heavy chain constant domain, and the light chain variable domain is attached to a single light chain variable domain, and the two constant domains associate together.
- the heavy and light chain variable domains are covalently linked together with a peptide, as in the single chain antibodies, or peptide sequences attached, preferably at the C-terminal end which will associate through forming cysteine bonds or through non-covalent interactions, such as the introduction of “leucine zipper” motifs.
- the Fv fragments are preferably used.
- variable domains isolated from a repertoire of variable region genes offer a way of building complete antibodies, and an alternative to hybridoma technology.
- complete antibodies may be made and should possess natural effector functions, such as complement lysis.
- This route is particularly attractive for the construction of human monoclonal antibodies, as hybridoma technology has proved difficult, and for example, although human peripheral blood lymphocytes can be immortalized with Epstein Barr virus, such hybridomas tend to secrete low affinity IgM antibodies.
- lymphocytes do not generally secrete antibodies directed against host-proteins.
- human antibodies directed against human proteins for example to human cell surface markers to treat cancers, or to histocompatibility antigens to treat auto-immune diseases.
- the construction of human antibodies built from the combinatorial repertoire of heavy and light chain variable domains may overcome this problem, as it will allow human antibodies to be built with specificities which would normally have been eliminated.
- the method also offers a new way of making bispecific antibodies.
- Antibodies with dual specificity can be made by fusing two hybridomas of different specificities, so as to make a hybrid antibody with an Fab arm of one specificity, and the other Fab arm of a second specificity.
- the yields of the bispecific antibody are low, as heavy and light chains also find the wrong partners.
- the construction of Fv fragments which are tightly associated should preferentially drive the association of the correct pairs of heavy with light chains. (It would not assist in the correct pairing of the two heavy chains with each other.)
- the improved production of bispecific antibodies would have a variety of applications in diagnosis and therapy, as is well known.
- the invention provides a species specific general oligonucleotide primer or a mixture of such primers useful for cloning variable domain encoding sequences from animals of that species.
- the method allows a single pair or pair of mixtures of species specific general primers to be used to clone any desired antibody specificity from that species. This eliminates the need to carry out any sequencing of the target sequence to be cloned and the need to design specific primers for each specificity to be recovered.
- variable genes for the expression of the variable genes directly on cloning, for the screening of the encoded domains for binding activities and for the assembly of the domains with other variable domains derived from the repertoire.
- mouse splenic ds mRNA or genomic DNA may be obtained from a hyper-immunized mouse.
- the expression vector would be used to transform a host cell, for instance a bacterial cell, to enable it to produce an Fv fragment or a Fab fragment.
- the Fv or Fab fragment would then be built into a monoclonal antibody by attaching constant domains and expressing it in mammalian cells.
- oligonucleotide primers or mixed primers were used. Their locations are marked on FIG. 1 and sequences are as follows:
- VH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCCAG 3′; VH1FOR-2 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCC 3′; Hu1VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu2VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu3VHFOR 5′ CTTGGTGGATGCTGAGGAGACGGTGACC 3′; Hu4VHFOR 5′ CTTGGTGGATGCTGATGAGACGGTGACC 3′; MOJH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTGCGCCCCCCAG 3′; MOJH2FOR 5′ TGAGGAGACGGTGACCGTGGTGCCTTGGCCCCAG 3′; MOJH3FOR 5′ TGCAGAGACGGTGACCAGTCCCTTGGCCCCAG 3′; MOJH4FOR 5′ TGAGGAGACGGTGACCGAGGTTCC
- VH1FOR is designed to anneal with the 3′ end of the sense strand of any mouse heavy chain variable domain encoding sequence. It contains a BstEII recognition site.
- VK1FOR is designed to anneal with the 3′ end of the sense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a BglII recognition site.
- VH1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse heavy chain variable domain and contains a PstI recognition site.
- VK1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a PvuII recognition site.
- MAbs monoclonal antibodies
- MBr1 BW431/26 [24]
- BW494/32 BW494/32 [25]
- BW250/183 [24,26] BW704/152 [27].
- MAb MBr1 is particularly interesting in that it is known to be specific for a saccharide epitope on a human mammary carcinoma line MCF-7 [28].
- a 50 ⁇ l reaction solution which contains 10 ⁇ g mRNA, 20 pmole VH1FOR primer, 250 ⁇ M each of dATP, dTTP, dCTP and dGTP, 10 mM dithiothreitol (DTT), 100 mM Tris.HCl, 10 MM MgCl 2 and 140 mM KCl, adjusted to pH 8.3 was prepared.
- the reaction solution was heated at 70° C. for ten minutes and allowed to cool to anneal the primer to the 3′ end of the variable domain encoding sequence in the mRNA.
- To the reaction solution was then added 46 units of reverse transcriptase (Anglian Biotec) and the solution was then incubated at 42° C. for 1 hour to cause first strand cDNA synthesis.
- variable domain encoding sequences were amplified as follows.
- a 50 ⁇ l reaction solution containing 5 ⁇ l of the ds RNA/DNA hybrid-containing solution, 25 pmole each of VH1FOR and VH1BACK primers, 250 ⁇ M of dATP, dTTP, dCTP and dGTP, 67 mM Tris.HCl, 17 mM ammonium sulphate, 10 mM MgCl 2 , 200 ⁇ g/ml gelatine and 2 units Taq polymerase (Cetus) was prepared.
- the reaction solution was overlaid with paraffin oil and subjected to 25 rounds of temperature cycling using a Techne PHC-1 programmable heating block. Each cycle consisted of 1 minute and 95° C. (to denature the nucleic acids), 1 minute at 30° C. (to anneal the primers to the nucleic acids) and 2 minutes at 72° C. (to cause elongation from the primers). After the 25 cycles, the reaction solution and the oil were extracted twice with ether, once with phenol and once with phenol/CHCl3. Thereafter ds cDNA was precipitated with ethanol. The precipitated ds cDNA was then taken up in 50 ⁇ l of water and frozen.
- VK1FOR and VK1BACK primers were used in place of the VH1FOR and VH1BACK primers respectively.
- a BstEII recognition site was introduced into the vector M13-HuVHNP [31] by site directed mutagenesis [32,33] to produce the vector M13-VHPCR1 ( FIGS. 2 and 3 ).
- Each amplified heavy chain variable domain encoding sequence was digested with the restriction enzymes PstI and BstEII.
- the fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VHPCR1 which had been digested with PstI and BstEII and purified on an 0.8% agarose gel.
- Clones containing the variable domain inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding variable gene in the M13-VHPCR1 vector.
- variable domain encoding sequences of BW431/26 There is an internal PstI site in the heavy chain variable domain encoding sequences of BW431/26. This variable domain encoding sequence was therefore assembled in two steps. The 3′ PstI-BstEII fragment was first cloned into M13-VHPCR1, followed in a second step by the 5′ PstI fragment.
- Vector M13 mp 18 [35] was cut with PvuII and the vector backbone was blunt ligated to a synthetic HindIII-BamHI polylinker.
- Vector M13-HuVKLYS [36] was digested with HindIII and BamHI to isolate the HuVKLYS gene. This HindIII-BamHI fragment was then inserted into the HindIII-BamHI polylinker site to form a vector M13-VKPCR1 which lacks any PvuII sites in the vector backbone ( FIGS. 4 and 5 ).
- This vector was prepared in E. coli JM110 [22] to avoid dam methylation at the BclI site.
- Each amplified light chain variable domain encoding sequence was digested with PvuII and BglII.
- the fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VKPCR1 which had been digested with PvuII and BclI, purified on an 0.8% agarose gel and treated with calf intestinal phosphatase.
- Clones containing the light chain variable region inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding region of the variable domain in the M13-VKPCR1 vector.
- nucleotide sequences of the MBr1 heavy and light chain variable domains are shown in FIG. 6 with part of the flanking regions of the M13-VHPCR1 and M13-VKPCR1 vectors.
- the HindIII-BamHI fragment carrying the MBr1 heavy chain variable domain encoding sequence in M13-VHPCR1 was recloned into a pSV-gpt vector with human ⁇ 1 constant regions [37] ( FIG. 7 ).
- the MBr1 light chain variable domain encoding sequence in M13-VKPCR1 was recloned as a HindIII-BamHI fragment into a pSV vector, PSV-hyg-HuCK with a hygromycin resistance marker and a human kappa constant domain ( FIG. 8 ).
- the assembly of the genes is summarized in FIG. 9 .
- the vectors thus produced were linearized with PvuI (in the case of the pSV-hygro vectors the PvuI digest is only partial) and cotransfected into the non-secreting mouse myeloma line NSO [38] by electroporation [39].
- PvuI in the case of the pSV-hygro vectors the PvuI digest is only partial
- NSO non-secreting mouse myeloma line NSO
- electroporation electroporation [39].
- One day after cotransfection cells were selected in 0.3 ⁇ g/ml mycophenolic acid (MPA) and after seven days in 1 ⁇ g/ml MPA. After 14 days, four wells, each containing one or two major colonies, were screened by incorporation of 14 C-lysine [40] and the secreted antibody detected after precipitation with protein-A SepharoseTM (Pharmacia) on SDS-PAGE [41].
- the gels were stained, fixed, soaked in
- the chimeric antibody in the supernatant like the parent mouse MBr1 antibody, was found to bind to MCF-7 cells but not the HT-29 cells, thus showing that the specificity had been properly cloned and expressed.
- the DNA from the mouse spleen was prepared in one of two ways (although other ways can be used).
- Method 1 A mouse spleen was cut into two pieces and each piece was put into a standard Eppendorf tube with 200 ⁇ l of PBS. The tip of a 1 ml glass pipette was closed and rounded in the blue flame of a Bunsen burner. The pipette was used to squash the spleen piece in each tube. The cells thus produced were transferred to a fresh Eppendorf tube and the method was repeated three times until the connective tissue of the spleen appeared white. Any connective tissue which has been transferred with the cells was removed using a drawn-out Pasteur pipette. The cells were then washed in PBS and distributed into four tubes.
- mice spleen cells were then sedimented by a 2 minute spin in a Microcentaur centrifuge at low speed setting. All the supernatant was aspirated with a drawn out Pasteur pipette. If desired, at this point the cell sample can be frozen and stored at ⁇ 20° C.
- the supernatant was transferred to a new tube and to this was added 125 ⁇ l 5M NaCl and 30 ⁇ l 1M MOPS adjusted to pH 7.0.
- the DNA in the supernatant was absorbed on a Quiagen 5 tip and purified following the manufacturer's instructions for lambda DNA. After isopropanol precipitation, the DNA was resuspended in 500 ⁇ l water.
- Method 2 This method is based on the technique described in Maniatis et al. [30].
- a mouse spleen was cut into very fine pieces and put into a 2 ml glass homogenizer. The cells were then freed from the tissue by several slow up and down strokes with the piston.
- the cell suspension was made in 500 ⁇ l phosphate buffered saline (PBS) and transferred to an Eppendorf tube. The cells were then spun for 2 min at low speed in a Microcentaur centrifuge. This results in a visible separation of white and red cells. The white cells, sedimenting slower, form a layer on top of the red cells. The supernatant was carefully removed and spun to ensure that all the white cells had sedimented.
- the layer of white cells was resuspended in two portions of 500 ⁇ l PBS and transferred to another tube.
- the white cells were precipitated by spinning in the Microcentaur centrifuge at low speed for one minute. The cells were washed a further two times with 500 ⁇ l PBS, and were finally resuspended in 200 ⁇ l PBS. The white cells were added to 2.5 ml 25 mM EDTA and 10 mM Tris.Cl, pH 7.4, and vortexed slowly. While vortexing 25 ⁇ l 20% SDS was added. The cells lysed immediately and the solution became viscous and clear. 100 ⁇ l of 20 mg/ml proteinase K was added and incubated one to three hours at 50° C.
- the sample was extracted with an equal volume of phenol and the same volume of chloroform, and vortexed. After centrifuging, the aqueous phase was removed and 1/10 volume 3M ammonium acetate was added. This was overlaid with three volumes of cold ethanol and the tube rocked carefully until the DNA strands became visible.
- the DNA was spooled out with a Pasteur pipette, the ethanol allowed to drip off, and the DNA transferred to 1 ml of 10 mM Tris.Cl pH 7.4, 0.1 mM EDTA in an Eppendorf tube. The DNA was allowed to dissolve in the cold overnight on a roller.
- the DNA solution was diluted 1/10 in water and boiled for 5 min prior to using the polymerase chain reaction (PCR).
- PCR polymerase chain reaction
- typically 50-200 ng of DNA were used.
- the heavy and light chain variable domain encoding sequences in the genomic DNA isolated from the human PBL or the mouse spleen cells was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA Hybrid” in Example 1, except that during the annealing part of each cycle, the temperature was held at 65° C. and that 30 cycles were used. Furthermore, to minimize the annealing between the 3′ ends of the two primers, the sample was first heated to 95° C., then annealed at 65° C., and only then was the Taq polymerase added. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- VH1FOR and VH1BACK The primers used to amplify the mouse spleen genomic DNA were VH1FOR and VH1BACK, for the heavy chain variable domain and VK2FOR and VK1BACK, for the light chain variable domain. (VK2FOR only differs from VK1FOR in that it has an extra C residue on the 5′ end.)
- VH1FOR Likewise mixtures of VH1FOR, MOJH1FOR, MOJH2FOR, MOJH3FOR and MOJH4FOR were used as forward primers and mixtures of VH1BACK, MOVHIBACK, MOVHIIABACK, MOVHIIBBACK, MOVHIIIBACK were used as backward primers for amplification of VH genes.
- All these heavy chain FOR primers referred to above contain a BstEII site and all the BACK primers referred to above contain a PstI site. These light chain FOR and BACK primers referred to above all contain BglII and PvuII sites respectively.
- Light chain primers VK3FOR and VK2BACK were also devised which utilized different restriction sites, SacI and XhoI.
- the preferred amplification conditions for mouse VH genes are as follows: the sample was made in a volume of 50-100 ⁇ l, 50-100 ng of DNA, VH1FOR-2 and VH1BACK primers (25 pmole of each), 250 ⁇ M of each deoxynucleotide triphosphate, 10 mM Tris.HCl, pH 8.8, 50 mM KCl, 1.5 mM MgCl 2 , and 100 ⁇ g/ml gelatine. The sample was overlaid with paraffin oil, heated to 95° C. for 2 min, 65° C.
- the preferred amplification conditions for mouse V ⁇ genes from genomic DNA are as follows: the sample treated as above except with V ⁇ primers, for example VK3FOR and VK2BACK, and using a cycle of 94° C. for one minute, 60° C. for one minute and 72° C. for one minute.
- V ⁇ primers for example VK3FOR and VK2BACK
- the conditions which were devised for genomic DNA are also suitable for amplification from the cDNA derived from mRNA from mouse spleen or mouse hybridoma.
- the reaction mixture was then extracted twice with 40 ⁇ l of water-saturated diethyl ether. This was followed by a standard phenol extraction and ethanol precipitation as described in Example 1.
- the DNA pellet was then dissolved in 100 ⁇ l 10 mM Tris.Cl, 0.1 mM EDTA.
- Each reaction mixture containing a light chain variable domain encoding sequence was digested with SacI and XhoI (or with PvuII and BglII) to enable it to be ligated into a suitable expression vector.
- Each reaction mixture containing a heavy chain variable domain encoding sequence was digested with PstI and BstEII for the same purpose.
- the heavy chain variable genes isolated as above from a mouse hyper-immunized with lysozyme were cloned into M13VHPCR1 vector and sequenced.
- the complete sequences of 48 VH gene clones were determined ( FIGS. 10 a - 10 b ). All but two of the mouse VH gene families were represented, with frequencies of: VA (1), IIIC (1), IIIB (8), IIIA (3), IIB (17), IIA (2), IB (12), IA (4).
- the D segments could be assigned to families SP2 (14), FL16 (11) and Q52 (5), and in 38 clones the JH minigenes to families JH1 (3), JH2 (7), JH3 (14) and JH4 (14).
- the different sequences of CDR3 marked out each of the 48 clones as unique. Nine pseudogenes and 16 unproductive rearrangements were identified. Of the clones sequenced, 27 have open reading frames.
- the method is capable of generating a diverse repertoire of heavy chain variable genes from mouse spleen DNA.
- Method 1 20 ml of heparinized human blood from a healthy volunteer was diluted with an equal volume of phosphate buffered saline (PBS) and distributed equally into 50 ml Falcon tubes. The blood was then underlayed with 15 ml Ficoll Hypaque (Pharmacia 10-A-001-07). To separate the lymphocytes from the red blood cells, the tubes were spun for 10 minutes at 1800 rpm at room temperature in an IEC Centra 3E table centrifuge. The peripheral blood lymphocytes (PBL) were then collected from the interphase by aspiration with a Pasteur pipette. The cells were diluted with an equal volume of PBS and spun again at 1500 rpm for 15 minutes. The supernatant was aspirated, the cell pellet was resuspended in 1 ml PBS and the cells were distributed into two Eppendorf tubes.
- PBS phosphate buffered saline
- Method 2 40 ml human blood from a patient with HIV in the pre-AIDS condition was layered on Ficoll to separate the white cells (see Method 1 above). The white cells were then incubated in tissue culture medium for 4-5 days. On day 3, they were infected with Epstein Barr virus. The cells were pelleted (approx 2 ⁇ 10 7 cells) and washed in PBS.
- the cells were pelleted again and lysed with 7 ml 5M guanidine isothiocyanate, 50 mM Tris, 10 mM EDTA, 0.1 mM dithiothreitol.
- the cells were vortexed vigorously and 7 volumes of 4M LiCl added.
- the mixture was incubated at 4° C. for 15-20 hrs.
- the suspension was spun and the supernatant resuspended in 3M LiCl and centrifuged again.
- the pellet was dissolved in 2 ml 0.1% SDS, 10 mM Tris HCl and 1 mM EDTA.
- the suspension was frozen at ⁇ 20° C., and thawed by vortexing for 20 s every 10 min for 45 min.
- RNA was precipitated by adding 1/10 volume 3M sodium acetate and 2 vol ethanol and leaving overnight at ⁇ 20° C. The pellet was suspended in 0.2 ml water and reprecipitated with ethanol. Aliquots for cDNA synthesis were taken from the ethanol precipitate which had been vortexed to create a fine suspension.
- the back primers for the amplification of human DNA were designed to match the available human heavy and light chain sequences, in which the different families have slightly different nucleotide sequences at the 5′ end.
- the primers Hu2VHIBACK, HuVHIIBACK, Hu2VHIIIBACK and HuVH1VBACK were designed as back primers, and HUJH1FOR, HUJH2FOR and HUJH4FOR as forward primers based entirely in the variable gene.
- Another set of forward primers Hu1VHFOR, Hu2VHFOR, Hu3VHFOR, and Hu4VHFOR was also used, which were designed to match the human J-regions and the 5′ end of the constant regions of different human isotypes.
- the amplified DNA from the separate primings was then pooled, digested with restriction enzymes PstI and BstEII as above, and then cloned into the vector M13VHPCR1 for sequencing.
- the sequences reveal a diverse repertoire ( FIG. 11 ) at this stage of the disease.
- HuJK1FOR, HUJK3FOR; HUJK4FOR and HUJK5FOR were used as forward primers and VK1BACK as back primer. Using these primers it was possible to see a band of amplified ds cDNA of the correct size by gel electrophoresis.
- Human peripheral blood lymphocytes of a patient with non-Hodgkins lymphoma were prepared as in Example 3 (Method 1).
- the genomic DNA was prepared from the PBL using the technique described in Example 2 (Method 2).
- the VH region in the isolated genomic DNA was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA hybrid” in Example 1 above, except that during the annealing part of each cycle, the temperature was held at 55° C. and that 30 cycles were used. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- the forward primer used was HuHep1FOR, which contains a SacI site. This primer is designed to anneal to the 3′ end of the unrearranged human VH region gene, and in particular includes a sequence complementary to the last three codons in the VH region gene and nine nucleotides downstream of these three codons.
- HuOcta1BACK, HuOcta2BACK and HuOcta3BACK was used as the back primer. These primers anneal to a sequence in the promoter region of the genomic DNA VH gene (see FIG. 1 ). 5 ⁇ l of the amplified DNA was checked on 2% agarose gels in TBE buffer and stained with ethidium bromide. A double band was seen of about 620 nucleotides which corresponds to the size expected for the unrearranged VH gene. The ds cDNA was digested with SacI and cloned into an M13 vector for sequencing. Although there are some sequences which are identical, a range of different unrearranged human VH genes were identified ( FIG. 12 ).
- VHLYS heavy chain variable domain
- D1.3 anti-lysozyme
- the heavy chain variable domain (VHLYS) of the D1.3 (anti-lysozyme) antibody was cloned into a vector similar to that described previously [42] but under the control of the lac z promoter, such that the VHLYS domain is attached to a pelB leader sequence for export into the periplasm.
- the vector was constructed by synthesis of the pelB leader sequence [43], using overlapping oligonucleotides, and cloning into a pUC 19 vector [35].
- the VHLYS domain of the D1.3 antibody was derived from a cDNA clone [44] and the construct (PSW1) sequenced ( FIG. 13 ).
- VKLYS light chain variable region
- the colonies were inoculated into 50 ml 2 ⁇ TY (with 1% glucose and 100 ⁇ g/ml ampicillin) and grown in flasks at 37° C. with shaking for 12-16 hr.
- the cells were centrifuged, the pellet washed twice with 50 mM sodium chloride, resuspended in 2 ⁇ TY medium containing 100 ⁇ g/ml ampicillin and the inducer IPTG (1 mM) and grown for a further 30 hrs at 37° C.
- the cells were centrifuged and the supernatant was passed through a Nalgene filter (0.45 ⁇ m) and then down a 1-5 ml lysozyme-Sepharose® affinity column (Pharmacia Fine Chemicals, Inc.).
- the column was derived by coupling lysozyme at 10 mg/ml to CNBr activated Sepharose.
- the column was first washed with phosphate buffered saline (PBS), then with 50 mM diethylamine to elute the VHLYS domain (from pSW1) or VHLYS in association with VKLYS (from pSW2).
- PBS phosphate buffered saline
- VHLYS and VKLYS domains were identified by SDS polyacrylamide electrophoresis as the correct size.
- N-terminal sequence determination of VHLYS and VKLYS isolated from a polyacrylamide gel showed that the signal peptide had been produced correctly.
- both the Fv fragment and the VHLYS domains are able to bind to the lysozyme affinity column, suggesting that both retain at least some of the affinity of the original antibody.
- VHLYS domain was compared by FPLC with that of the Fv fragment on Superose 12. This indicates that the VHLYS domain is a monomer.
- the binding of the VHLYS and Fv fragment to lysozyme was checked by ELISA, and equilibrium and rapid reaction studies were carried out using fluorescence quench.
- the ELISA for lysozyme binding was undertaken as follows:
- VHLYS or Fv fragment VHLYS associated with VKLYS
- the reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid pH 4.3.
- ELISA plates were read in a Titertek Multiscan plate reader. Supernatant from the induced bacterial cultures of both pSW1 (VHLYS domain) or pSW2 (Fv fragment) was found to bind to lysozyme in the ELISA.
- the purified VHLYS and Fv fragments were titrated with lysozyme using fluorescence quench (Perkin Elmer LS5B Luminescence Spectrometer) to measure the stoichiometry of binding and the affinity constant for lysozyme [48,49].
- fluorescence quench Perkin Elmer LS5B Luminescence Spectrometer
- the titration of the Fv fragment at a concentration of 30 nM indicates a dissociation constant of 2.8 nM using a Scatchard analysis.
- VHLYS was titrated with lysozyme as above using fluorescence quench.
- the on-rates for VHLYS and Fv fragments with lysozyme were determined by stopped-flow analysis (HI Tech Stop Flow SHU machine) under pseudo-first order conditions with the fragment at a ten fold higher concentration than lysozyme [50].
- the concentration of lysozyme binding sites was first measured by titration with lysozyme using fluorescence quench as above. The on rates were calculated per mole of binding site (rather than amount of VHLYS protein).
- the on-rate for the Fv fragment was found to be 2.2 ⁇ 10 6 M ⁇ 1 s ⁇ 1 at 25° C.
- the on-rate for the VHLYS fragment found to be 3.8 ⁇ 10 6 M ⁇ 1 s ⁇ 1 and the off-rate 0.075 s ⁇ 1 at 20° C.
- the calculated affinity constant is 19 nM.
- the VHLYS binds to lysozyme with a dissociation constant of about 19 nM, compared with that of the Fv of 3 nM.
- a mouse was immunized with hen egg white lysozyme (100 ⁇ g i.p. day 1 in complete Freunds adjuvant), after 14 days immunized i.p. again with 100 ⁇ g lysozyme with incomplete Freunds adjuvant, and on day 35 i.v. with 50 ⁇ g lysozyme in saline. On day 39, spleen was harvested. A second mouse was immunized with keyhole limpet hemocyanin (KLH) in a similar way. The DNA was prepared from the spleen according to Example 2 (Method 2). The VH genes were amplified according to the preferred method in Example 2.
- KLH keyhole limpet hemocyanin
- Human peripheral blood lymphocytes from a patient infected with HIV were prepared as in Example 3 (Method 2) and mRNA prepared.
- the VH genes were amplified according to the method described in Example 3, using primers designed for human VH gene families.
- the reaction mixture and oil were extracted twice with ether, once with phenol and once with phenol/CHCl 3 .
- the double stranded DNA was then taken up in 50 ⁇ l of water and frozen. 5 ⁇ l was digested with PstI and BstEII (encoded within the amplification primers) and loaded on an agarose gel for electrophoresis. The band of amplified DNA at about 350 bp was extracted.
- the repertoire of amplified heavy chain variable domains was then cloned directly into the expression vector pSW1HPOLYMYC.
- This vector is derived from pSW1 except that the VHLYS gene has been removed and replaced by a polylinker restriction site.
- a sequence encoding a peptide tag was inserted ( FIG. 15 ). Colonies were toothpicked into 1 ml cultures. After induction (see Example 5 for details), 10 ⁇ l of the supernatant from fourteen 1 ml cultures was loaded on SDS-PAGE gels and the proteins transferred electrophoretically to nitrocellulose. The blot was probed with antibody 9E10 directed against the peptide tag.
- the probing was undertaken as follows.
- the nitrocellulose filter was incubated in 3% bovine serum albumin (BSA)/TBS buffer for 20 min (10 ⁇ TBS buffer is 100 mM Tris.HCl, pH 7.4, 9% w/v NaCl).
- BSA bovine serum albumin
- the filter was incubated in a suitable dilution of antibody 9E10 (about 1/500) in 3% BSA/TBS for 1-4 hrs. After three washes in TBS (100 ml per wash, each wash for 10 min), the filter was incubated with 1:500 dilution of anti-mouse antibody (peroxidase conjugated anti-mouse Ig (Dakopats)) in 3% BSA/TBS for 1-2 hrs.
- anti-mouse antibody peroxidase conjugated anti-mouse Ig (Dakopats)
- Colonies were then toothpicked individually into wells of an ELISA plate (200 ⁇ l) for growth and induction. They were assayed for lysozyme binding with the 9E10 antibody (as in Examples 5 and 7). Wells with lysozyme-binding activity were identified. Two positive wells (of 200) were identified from the amplified mouse spleen DNA and one well from the human cDNA. The heavy chain variable domains were purified on a column of lysozyme-Sepharose. The affinity for lysozyme of the clones was estimated by fluorescence quench titration as >50 nM.
- VH3 and VH8 The affinities of the two clones (VH3 and VH8) derived from the mouse genes were also estimated by stop flow analysis (ratio of k off /k on ) as 12 nM and 27 nM respectively. Thus both these clones have a comparable affinity to the VHLYS domain.
- the encoded amino acid sequences of VH3 and VH8 are given in FIG. 16 , and that of the human variable domain in FIG. 17 .
- a library of VH domains made from the mouse immunized with lysozyme was screened for both lysozyme and keyhole limpet hemocyanin (KLH) binding activities. Two thousand colonies were toothpicked in groups of five into wells of ELISA plates, and the supernatants tested for binding to lysozyme coated plates and separately to KLH coated plates. Twenty one supernatants were shown to have lysozyme binding activities and two to have KLH binding activities.
- a second expression library, prepared from a mouse immunized with KLH was screened as above. Fourteen supernatants had KLH binding activities and a single supernatant had lysozyme binding activity.
- a single rearranged VH gene it may be possible to derive entirely new antigen binding activities by extensively mutating each of the CDRs.
- the mutagenesis might be entirely random, or be derived from pre-existing repertoires of CDRs.
- a repertoire of CDR3s might be prepared as in the preceding examples by using “universal” primers based in the flanking sequences, and likewise repertoires of the other CDRs (singly or in combination).
- the CDR repertoires could be stitched into place in the flanking framework regions by a variety of recombinant DNA techniques.
- CDR3 appears to be the most promising region for mutagenesis as CDR3 is more variable in size and sequence than CDRs 1 and 2. This region would be expected to make a major contribution to antigen binding.
- the heavy chain variable region (VHLYS) of the anti-lysozyme antibody D1.3 is known to make several important contacts in the CDR3 region.
- the source of the heavy chain variable domain was an M113 vector containing the VHLYS gene.
- the body of the sequence encoding the variable region was amplified using the polymerase chain reaction (PCR) with the mutagenic primer VHMUT1 based in CDR3 and the M13 primer which is based in the M13 vector backbone.
- the mutagenic primer hypermutates the central four residues of CDR3 (Arg-Asp-Tyr-Arg).
- the PCR was carried out for 25 cycles on a Techne PHC-1 programmable heat block using 100 ng single stranded M13 mp19SWO template, with 25 pmol of VHMUT1 and the M13 primer, 0.5 mM each dNTP, 67 mM Tris.HCl, pH 8.8, 10 mM MgCl 2 , 17 mM (NH 4 ) 2 SO 4 , 200 ⁇ g/ml gelatine and 2.5 units Taq polymerase in a final volume of 50 ⁇ l.
- the temperature regime was 95° C. for 1.5 min, 25° C. for 1.5 min and 72° C. for 3 min (However a range of PCR conditions could be used).
- the reaction products were extracted with phenol/chloroform, precipitated with ethanol and resuspended in 10 mM Tris. HCl and 0.1 mM EDTA, pH 8.0.
- the products from the PCR were digested with PstI and BstEII and purified on a 1.5% LGT agarose gel in Tris acetate buffer using Geneclean® (Bio 101, LaJolla).
- the gel purified band was ligated into pSW2HPOLY ( FIG. 19 ).
- the vector was first digested with BstEII and PstI and treated with calf-intestinal phosphatase. Aliquots of the reaction mix were used to transform E. coli BMH 71-18 to ampicillin resistance. Colonies were selected on ampicillin (100 ⁇ g/ml) rich plates containing glucose at 0.8% w/v.
- Colonies resulting from transfection were picked in pools of five into two 96 well Corning microtitre plates, containing 200 ⁇ l 2 ⁇ TY medium and 100 ⁇ l TY medium, 100 ⁇ g/ml ampicillin and 1% glucose. The colonies were grown for 24 hours at 37° C. and then cells were washed twice in 200 ⁇ l 50 mM NaCl, pelleting the cells in an IEC Centra-3 bench top centrifuge with microtitre plate head fitting. Plates were spun at 2,500 rpm for 10 min at room temperature. Cells were resuspended in 200 ⁇ l 2 ⁇ TY, 100 ⁇ g/ml ampicillin and 1 mM IPTG (Sigma) to induce expression, and grown for a further 24 hr.
- plasmids were prepared and the VKLYS gene excised by cutting with EcoRI and religating. Thus the plasmids should only direct the expression of the VHLYS mutants. 1.5 ml cultures were grown and induced for expression as above. The cells were spun down and supernatant shown to bind lysozyme as above. (Alternatively the amplified mutant VKLYS genes could have been cloned directly into the pSW1HPOLY vector for expression of the mutant activities in the absence of VKLYS.)
- An ELISA method was devised in which the activities of bacterial supernatants for binding of lysozyme (or KLH) were compared.
- a vector was devised for tagging of the VH domains at its C-terminal region with a peptide from the c-myc protein which is recognized by a monoclonal antibody 9E10.
- the vector was derived from pSW1 by a BstEII and SmaI double digest, and ligation of an oligonucleotide duplex made from
- VHLYSMYC protein domain expressed after induction was shown to bind to lysozyme and to the 9E10 antibody by ELISA as follows:
- test solution was discarded, and the wells washed out with PBS (3 washes).
- 100 ⁇ l of 4 ⁇ g/ml purified 9E10 antibody in 2% Sainsbury's instant dried skimmed milk powder in PBS was added, and incubated at 37° C. for 2 hrs;
- reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid, pH 4.3.
- ELISA plates were read in an Titertek Multiscan plate reader.
- VHLYSMUT59 To check the affinity of the VHLYSMUT59 domain directly, the clone was grown at the 1 L scale and 200-300 ⁇ g purified on lysozyme-Sepharose as in Example 5. By fluorescence quench titration of samples of VHLYS and VHLYSMUT59, the number of binding sites for lysozyme were determined. The samples of VHLYS and VHLYSMUT59 were then compared in the competition ELISA with VHLYSMYC over two orders of magnitude. In the competition assay each microtitre well contained a constant amount of VHLYSMYC (approximately 0.6 ⁇ g VHLYSMYC).
- VHLYS or VHLYSMUT59 Varying amounts of VHLYS or VHLYSMUT59 (3.8 ⁇ M in lysozyme binding sites) were added (0.166-25 ⁇ l). The final volume and buffer concentration in all wells was constant. 9E10 (anti-myc) antibody was used to quantitate bound VHLYSMYC in each assay well. The % inhibition of VHLYSMYC binding was calculated for each addition of VHLYS or VHLYSMUT59, after subtraction of background binding. Assays were carried out in duplicate. The results indicate that VHLYSMUT59 has a higher affinity for lysozyme than VHLYS.
- VHLYSMUT59 gene was sequenced (after recloning into M13) and shown to be identical to the VHLYS gene except for the central residues of CDR3 (Arg-Asp-Tyr-Arg). These were replaced by Thr-Gln-Arg-Pro: (encoded by ACACAAAGGCCA).
- a library of 2000 mutant VH clones was screened for lysozyme and also for KLH binding (toothpicking 5 colonies per well as described in Example 6).
- Nineteen supernatants were identified with lysozyme binding activities and four with KLH binding activities. This indicates that new specificities and improved affinities can be derived by making a random repertoire of CDR3.
- Two copies of the cloned heavy chain variable gene of the D1.3 antibody were linked by a nucleotide sequence encoding a flexible linker Gly-Gly-Gly-Ala-Pro-Ala-Ala-Ala-Pro-Ala-Gly-Gly-Gly- (by several steps of cutting, pasting and site directed mutagenesis) to yield the plasmid pSW3 ( FIG. 20 ).
- the expression was driven by a lacZ promoter and the protein was secreted into the periplasm via a pelB leader sequence (as described in Example 5 for expression of pSW1 and pSW2).
- the protein could be purified to homogeneity on a lysozyme affinity column.
- a cysteine residue was introduced at the C-terminus of the VHLYS domain in the vector pSW2.
- the cysteine was introduced by cleavage of the vector with the restriction enzymes BstI and SmaI (which excises the C-terminal portion of the J segment) and ligation of a short oligonucleotide duplex
- FIGS. 21 a - 21 c there is shown the sequence of a fusion of a VH domain with alkaline phosphatase.
- the alkaline phosphatase gene was cloned from a plasmid carrying the E. coli alkaline phosphatase gene in a plasmid pEK48 [51] using the polymerase chain reaction. The gene was amplified with the primers
- FIGS. 21 a - 21 c The construction ( FIGS. 21 a - 21 c ) was expressed in E.
- Example 5 coli strain BMH71-18 as in Example 5 and screened for phosphatase activity using 1 mg/ml p-nitrophenylphosphate as substrate in 10 mM diethanolamine and 0.5 mM MgCl 2 , pH 9.5) and also on SDS polyacrylamide gels which had been Western blotted (detecting with anti-idiotypic antiserum). No evidence was found for the secretion of the linked VHLYS-alkaline phosphatase as detected by Western blots (see Example 5), or for secretion of phosphatase activity.
- linker sequences could then be introduced at the BstEII site to improve the spacing between the two domains.
- V ⁇ genes A repertoire of V ⁇ genes was derived by PCR using primers as described in Example 2 from DNA prepared from mouse spleen and also from mouse spleen mRNA using the primers VK3FOR and VK2BACK and a cycle of 94° C. for 1 min, 60° C. for 1 min, 72° C. for 2 min.
- the PCR amplified DNA was fractionated on the agarose gel, the band excised and cloned into a vector which carries the VHLYS domain (from the D 1.3 antibody), and a cloning site (SacI and XhoI) for cloning of the light chain variable domains with a myc tail (pSW1VHLYS-VKPOLYMYC, FIG. 22 ).
- Clones were screened for lysozyme binding activities as described in Examples 5 and 7 via the myc tag on the light chain variable domain, as this should permit the following kinds of V ⁇ domains to be identified:
- VHLYS domain was replaced by the heavy chain variable domain VH3 which had been isolated from the repertoire (see Example 6), and then the V ⁇ domains cloned into the vector. (Note that the VH3 domain has an internal SacI site and this was first removed to allow the cloning of the V ⁇ repertoire as SacI-XhoI fragments.)
- the present invention enables the cloning, amplification and expression of heavy and light chain variable domain encoding sequences in a much more simple manner than was previously possible. It also shows that isolated variable domains or such domains linked to effector molecules are unexpectedly useful.
Abstract
Description
- This is a continuation of application Ser. No. 10/290,233, filed Nov. 8, 2002 (allowed), which is a continuation of application Ser. No. 09/722,364, filed Nov. 28, 2000 (now U.S. Pat. No. 6,545,142), which is a continuation of application Ser. No. 08/470,031, filed Jun. 6, 1995 (now U.S. Pat. No. 6,248,516), which is a divisional of application Ser. No. 08/332,046, filed Nov. 1, 1994 (now abandoned), which is a continuation of application Ser. No. 07/796,805, filed Nov. 25, 1991 (now abandoned), which is a divisional of application Ser. No. 07/580,374, filed Sep. 11, 1990 (now abandoned), which is a continuation of PCT Application No. PCT/GB89/01344, filed Nov. 13, 1989, the entire contents of each of which is hereby incorporated by reference in this application.
- The present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- The present invention is now described, by way of example only, with reference to the accompanying drawings.
-
FIG. 1 shows a schematic representation of the unrearranged and rearranged heavy and light chain variable genes and the location of the primers. -
FIG. 2 shows a schematic representation of the M13-VHPCR1 vector and a cloning scheme for amplified heavy chain variable domains. -
FIG. 3 shows the sequence of the Ig variable region derived sequences in M13-VHPCR1. -
FIG. 4 shows a schematic representation of the M13-VKPCR1 vector and a cloning scheme for light chain variable domains. -
FIG. 5 shows the sequence of the Ig variable region derived sequences in M13-VKPCR1. -
FIG. 6 shows the nucleotide sequences of the heavy and light chain variable domain encoding sequences of MAb MBr1. -
FIG. 7 shows a schematic representation of the pSV-gpt vector (also known as α-Lys 30) which contains a variable region cloned as a HindIII-BamHI fragment, which is excised on introducing the new variable region. The gene for human IgG1 has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique. -
FIG. 8 shows a schematic representation of the pSV-hygro vector (also known as α-Lys 17). It is derived from pSV gpt vector with the gene encoding mycophenolic acid replaced by a gene coding for hygromycin resistance. The construct contains a variable gene cloned as a HindIII-BamHI fragment which is excised on introducing the new variable region. The gene for human Cκ has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique. -
FIG. 9 shows the assembly of the mouse: human MBr1 chimeric antibody. -
FIGS. 10 a-10 b shows encoded amino acid sequences of 48 mouse rearranged VH genes. -
FIG. 11 shows encoded amino acid sequences of human rearranged VH genes. -
FIG. 12 shows encoded amino acid sequences of unrearranged human VH genes. -
FIG. 13 shows the sequence of part of the plasmid pSW1: essentially the sequence of a pectate lyase leader linked to VHLYS in pSW1 and cloned as an SphI-EcoRI fragment into pUC19 and the translation of the open reading frame encoding the pectate lyase leader-VHLYS polypeptide being shown. -
FIGS. 14 a-14 b shows the sequence of part of the plasmid pSW2: essentially the sequence of a pectate lyase leader linked to VHLYS and to VKLYS, and cloned as an SphI-EcoRI-EcoRI fragment into pUC19 and the translation of open reading frames encoding the pectate lyase leader-VHLYS and pectate lyase leader-VKLYS polypeptides being shown. -
FIG. 15 shows the sequence of part of the plasmid pSW1HPOLYMYC which is based on pSW1 and in which a polylinker sequence has replaced the variable domain of VHLYS, and acts as a cloning site for amplified VH genes, and a peptide tag is introduced at the C-terminal end. -
FIG. 16 shows the encoded amino acid sequences of two VH domains derived from mouse spleen and having lysozyme binding activity, and compared with the VH domain of the D1,3 antibody. The arrows mark the points of difference between the two VH domains. -
FIG. 17 shows the encoded amino acid sequence of a VH domain derived from human peripheral blood lymphocytes and having lysozyme binding activity. -
FIG. 18 shows a scheme for generating and cloning mutants of the VHLYS gene, which is compared with the scheme for cloning natural repertoires of VH genes. -
FIG. 19 shows the sequence of part of the vector pSW2HPOLY. -
FIG. 20 shows the sequence of part of the vector pSW3 which encodes the two linked VHLYS domains. -
FIGS. 21 a-21 c shows the sequence of the VHLYS domain and pelB leader sequence fused to the alkaline phosphatase gene. -
FIG. 22 shows the sequence of the vector pSW1VHLYS-VKPOLYMYC for expression of a repertoire of Vκ light chain variable domains in association with the VHLYS domain. -
FIG. 23 shows the sequence of VH domain which is secreted at high levels from E. coli. The differences with VHLYS domain are marked. - The present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- A list of references is appended to the end of the description. The documents listed therein are referred to in the description by number, which is given in square brackets.
- The Ig superfamily includes not only the Igs themselves but also such molecules as receptors on lymphoid cells such as T lymphocytes. Immunoglobulins comprise at least one heavy and one light chain covalently bonded together. Each chain is divided into a number of domains. At the N-terminal end of each chain is a variable domain. The variable domains on the heavy and light chains fit together to form a binding site designed to receive a particular target molecule. In the case of Igs, the target molecules are antigens. T-cell receptors have two chains of equal size, the α and β chains, each consisting of two domains. At the N-terminal end of each chain is a variable domain and the variable domains on the α and β chains are believed to fit together to form a binding site for target molecules, in this case peptides presented by a histocompatibility antigen. The variable domains are so called because their amino acid sequences vary particularly from one molecule to another. This variation in sequence enables the molecules to recognize an extremely wide variety of target molecules.
- Much research has been carried out on Ig molecules to determine how the variable domains are produced. It has been shown that each variable domain comprises a number of areas of relatively conserved sequence and three areas of hypervariable sequence. The three hypervariable areas are generally known as complementarity determining regions (CDRs).
- Crystallographic studies have shown that in each variable domain of an Ig molecule the CDRs are supported on framework areas formed by the areas of conserved sequences. The three CDRs are brought together by the framework areas and, together with the CDRs on the other chain, form a pocket in which the target molecule is received.
- Since the advent of recombinant DNA technology, there has been much interest in the use of such technology to clone and express Ig molecules and derivatives thereof. This interest is reflected in the numbers of patent applications and other publications on the subject.
- The earliest work on the cloning and expression of full Igs in the patent literature is EP-
A-0 120 694 (Boss). The Boss application also relates to the cloning and expression of chimeric antibodies. Chimeric antibodies are Ig-type molecules in which the variable domains from one Ig are fused to constant domains from another Ig. Usually, the variable domains are derived from an Ig from one species (often a mouse Ig) and the constant domains are derived from an Ig from a different species (often a human Ig). - A later European patent application, EP-A-0 125 023 (Genentech), relates to much the same subject as the Boss application, but also relates to the production by recombinant DNA technology of other variations of Ig-type molecules.
- EP-
A-0 194 276 (Neuberger) discloses not only chimeric antibodies of the type disclosed in the Boss application but also chimeric antibodies in which some or all of the constant domains have been replaced by non-Ig derived protein sequences. For instance, the heavy chain CH2 and CH3 domains may be replaced by protein sequences derived from an enzyme or a protein toxin. - EP-A-0 239 400 (Winter) discloses a different approach to the production of Ig molecules. In this approach, only the CDRs from a first type of Ig are grafted onto a second type of Ig in place of its normal CDRs. The Ig molecule thus produced is predominantly of the second type, since the CDRs form a relatively small part of the whole Ig. However, since the CDRs are the parts which define the specificity of the Ig, the Ig molecule thus produced has its specificity derived from the first Ig.
- Hereinafter, chimeric antibodies, CDR-grafted Igs, the altered antibodies described by Genentech, and fragments of such Igs such as F(ab′)2 and Fv fragments are referred to herein as modified antibodies.
- One of the main reasons for all the activity in the Ig field using recombinant DNA technology is the desire to use Igs in therapy. It is well known that, using the hybridoma technique developed by Kohler and Milstein, it is possible to produce monoclonal antibodies (MAbs) of almost any specificity. Thus, MAbs directed against cancer antigens have been produced. It is envisaged that these MAbs could be covalently attached or fused to toxins to provide “magic bullets” for use in cancer therapy. MAbs directed against normal tissue or cell surface antigens have also been produced. Labels can be attached to these so that they can be used for in vivo imaging.
- The major obstacle to the use of such MAbs in therapy or in vivo diagnosis is that the vast majority of MAbs which are produced are of rodent, in particular mouse, origin. It is very difficult to produce human MAbs. Since most MAbs are derived from non-human species, they are antigenic in humans. Thus, administration of these MAbs to humans generally results in an anti-Ig response being mounted by the human. Such a response can interfere with therapy or diagnosis, for instance by destroying or clearing the antibody quickly, or can cause allergic reactions or immune complex hypersensitivity which has adverse effects on the patient.
- The production of modified Igs has been proposed to ensure that the Ig administered to a patient is as “human” as possible, but still retains the appropriate specificity. It is therefore expected that modified Igs will be as effective as the MAb from which the specificity is derived but at the same time not very antigenic. Thus, it should be possible to use the modified Ig a reasonable number of times in a treatment or diagnosis regime.
- At the level of the gene, it is known that heavy chain variable domains are encoded by a “rearranged” gene which is built from three gene segments: an “unrearranged” VH gene (encoding the N-terminal three framework regions, first two complete CDRs and the first part of the third CDR), a diversity (DH)-segment (DH) (encoding the central portion of the third CDR) and a joining segment (JH) (encoding the last part of the third CDR and the fourth framework region). In the maturation of B-cells, the genes rearrange so that each unrearranged VH gene is linked to one DH gene and one JH gene. The rearranged gene corresponds to VH-DH-JH. This rearranged gene is linked to a gene which encodes the constant portion of the Ig chain.
- For light chains, the situation is similar, except that for light chains there is no diversity region. Thus light chain variable domains are encoded by an “unrearranged” VL gene and a JL gene. There are two types of light chains, kappa (κ) or lambda (λ), which are built respectively from unrearranged Vκ genes and Jκ segments, and from unrearranged Vλ genes and Jλ segments.
- Previous work has shown that it is necessary to have two variable domains in association together for efficient binding. For example, the associated heavy and light chain variable domains were shown to contain the antigen binding site [1]. This assumption is borne out by X-ray crystallographic studies of crystallized antibody/antigen complexes [2-6] which show that both the heavy and light chains of the antibody's variable domains contact the antigen. The expectation that association of heavy and light chain variable domains is necessary for efficient antigen binding underlies work to co-secrete these domains from bacteria [1], and to link the domains together by a short section of polypeptide as in the single chain antibodies [8, 9].
- Binding of isolated heavy and light chains had also been detected. However the evidence suggested strongly that this was a property of heavy or light chain dimmers. Early work, mainly with polyclonal antibodies, in which antibody heavy and light chains had been separated under denaturing conditions [10] suggested that isolated antibody heavy chains could bind to protein antigens [11] or hapten [12]. The binding of protein antigen was not characterized, but the hapten-binding affinity of the heavy chain fragments was reduced by two orders of magnitude [12] and the number of hapten molecules binding were variously estimated as 0.14 or 0.37 [13] or 0.26 [14] per isolated heavy chain. Furthermore binding of haptens was shown to be a property of dimeric heavy or dimeric light chains [14]. Indeed light chain dimers have been crystallized. It has been shown that in light chain dimers the two chains form a cavity which is able to bind to a single molecule of hapten [15].
- This confirms the assumption that, in order to obtain efficient binding, it is necessary to have a dimer, and preferably a heavy chain/light chain dimer, containing the respective variable domains. This assumption also underlies the teaching of the patent references cited above, wherein the intention is always to produce dimeric, and preferably heavy/light chain dimeric, molecules.
- It has now been discovered, contrary to expectations, that isolated Ig heavy chain variable domains can bind to antigen in a 1:1 ratio and with binding constants of equivalent magnitude to those of complete antibody molecules. In view of what was known up until now and in view of the assumptions made by those skilled in the art, this is highly surprising.
- Therefore, according to a first aspect of the present invention, there is provided a single domain ligand consisting of at least part of the variable domain of one chain of a molecule from the Ig superfamily.
- Preferably, the ligand consists of the variable domain of an Ig light, or, most preferably, heavy chain.
- The ligand may be produced by any known technique, for instance by controlled cleavage of Ig superfamily molecules or by peptide synthesis. However, preferably the ligand is produced by recombinant DNA technology. For instance, the gene encoding the rearranged gene for a heavy chain variable domain may be produced, for instance by cloning or gene synthesis, and placed into a suitable expression vector. The expression vector is then used to transform a compatible host cell which is then cultured to allow the ligand to be expressed and, preferably, secreted.
- If desired, the gene for the ligand can be mutated to improve the properties of the expressed domain, for example to increase the yields of expression or the solubility of the ligand, to enable the ligand to bind better, or to introduce a second site for covalent attachment (by introducing chemically reactive residues such as cysteine and histidine) or non-covalent binding of other molecules. In particular it would be desirable to introduce a second site for binding to serum components, to prolong the residence time of the domains in the serum; or for binding to molecules with effector functions, such as components of complement, or receptors on the surfaces of cells.
- Thus, hydrophobic residues which would normally be at the interface of the heavy chain variable domain with the light chain variable domain could be mutated to more hydrophilic residues to improve solubility; residues in the CDR loops could be mutated to improve antigen binding; residues on the other loops or parts of the β-sheet could be mutated to introduce new binding activities. Mutations could include single point mutations, multiple point mutations or more extensive changes and could be introduced by any of a variety of recombinant DNA methods, for example gene synthesis, site directed mutagenesis or the polymerase chain reaction.
- Since the ligands of the present invention have equivalent binding affinity to that of complete Ig molecules, the ligands can be used in many of the ways as are Ig molecules or fragments. For example, Ig molecules have been used in therapy (such as in treating cancer, bacterial and viral diseases), in diagnosis (such as pregnancy testing), in vaccination (such as in producing anti-idiotypic antibodies which mimic antigens), in modulation of activities of hormones or growth factors, in detection, in biosensors and in catalysis.
- It is envisaged that the small size of the ligands of the present invention may confer some advantages over complete antibodies, for example, in neutralizing the activity of low molecular weight drugs (such as dioxin) and allowing their filtration from the kidneys with drug attached; in penetrating tissues and tumors; in neutralizing viruses by binding to small conserved regions on the surfaces of viruses such as the “canyon” sites of viruses [16]; in high resolution epitope mapping of proteins; and in vaccination by ligands which mimic antigens.
- The present invention also provides receptors comprising a ligand according to the first aspect of the invention linked to one or more of an effector molecule, a label, a surface, or one or more other ligands having the same or different specificity.
- A receptor comprising a ligand linked to an effector molecule may be of use in therapy. The effector molecule may be a toxin, such as ricin or pseudomonas exotoxin, an enzyme which is able to activate a prodrug, a binding partner or a radio-isotope. The radio-isotope may be directly linked to the ligand or may be attached thereto by a chelating structure which is directly linked to the ligand. Such ligands with attached isotopes are much smaller than those based on Fv fragments, and could penetrate tissues and access tumors more readily.
- A receptor comprising a ligand linked to a label may be of use in diagnosis. The label may be a heavy metal atom or a radio-isotope, in which case the receptor can be used for in vivo imaging using X-ray or other scanning apparatus. The metal atom or radio-isotope may be attached to the ligand either directly or via a chelating structure directly linked to the ligand. For in vitro diagnostic testing, the label may be a heavy metal atom, a radio-isotope, an enzyme, a fluorescent or colored molecule or a protein or peptide tag which can be detected by an antibody, an antibody fragment or another protein. Such receptors would be used in any of the known diagnostic tests, such as ELISA or fluorescence-linked assays.
- A receptor comprising a ligand linked to a surface, such as a chromatography medium, could be used for purification of other molecules by affinity chromatography. Linking of ligands to cells, for example to the outer membrane proteins of E. coli or to hydrophobic tails which localize the ligands in the cell membranes, could allow a simple diagnostic test in which the bacteria or cells would agglutinate in the presence of molecules bearing multiple sites for binding the ligand(s).
- Receptors comprising at least two ligands can be used, for instance, in diagnostic tests. The first ligand will bind to a test antigen and the second ligand will bind to a reporter molecule, such as an enzyme, a fluorescent dye, a colored dye, a radio-isotope or a colored-, fluorescently- or radio-labelled protein.
- Alternatively, such receptors may be useful in increasing the binding to an antigen. The first ligand will bind to a first epitope of the antigen and the second ligand will bind to a second epitope. Such receptors may also be used for increasing the affinity and specificity of binding to different antigens in close proximity on the surface of cells. The first ligand will bind to the first antigen and the second epitope to the second antigen: strong binding will depend on the co-expression of the epitopes on the surface of the cell. This may be useful in therapy of tumors, which can have elevated expression of several surface markers. Further ligands could be added to further improve binding or specificity. Moreover, the use of strings of ligands, with the same or multiple specificities, creates a larger molecule which is less readily filtered from the circulation by the kidney.
- For vaccination with ligands which mimic antigens, the use of strings of ligands may prove more effective than single ligands, due to repetition of the immunizing epitopes.
- If desired, such receptors with multiple ligands could include effector molecules or labels so that they can be used in therapy or diagnosis as described above.
- The ligand may be linked to the other part of the receptor by any suitable means, for instance by covalent or non-covalent chemical linkages. However, where the receptor comprises a ligand and another protein molecule, it is preferred that they are produced by recombinant DNA technology as a fusion product. If necessary, a linker peptide sequence can be placed between the ligand and the other protein molecule to provide flexibility.
- The basic techniques for manipulating Ig molecules by recombinant DNA technology are described in the patent references cited above. These may be adapted in order to allow for the production of ligands and receptors according to the invention by means of recombinant DNA technology.
- Preferably, where the ligand is to be used for in vivo diagnosis or therapy in humans, it is humanized, for instance by CDR replacement as described in EP-A-0 239 400.
- In order to obtain a DNA sequence encoding a ligand, it is generally necessary firstly to produce a hybridoma which secretes an appropriate MAb. This can be a very time consuming method. Once an immunized animal has been produced, it is necessary to fuse separated spleen cells with a suitable myeloma cell line, grow up the cell lines thus produced, select appropriate lines, reclone the selected lines and reselect. This can take some long time. This problem also applies to the production of modified Igs.
- A further problem with the production of ligands, and also receptors according to the invention and modified Igs, by recombinant DNA technology is the cloning of the variable domain encoding sequences from the hybridoma which produces the MAb from which the specificity is to be derived. This can be a relatively long method involving the production of a suitable probe, construction of a clone library from cDNA or genomic DNA, extensive probing of the clone library, and manipulation of any isolated clones to enable the cloning into a suitable expression vector. Due to the inherent variability of the DNA sequences encoding Ig variable domains, it has not previously been possible to avoid such time consuming work. It is therefore a further aim of the present invention to provide a method which enables substantially any sequence encoding an Ig superfamily molecule variable domain (ligand) to be cloned in a reasonable period of time.
- According to another aspect of the present invention therefore, there is provided a method of cloning a sequence (the target sequence) which encodes at least part of the variable domain of an Ig superfamily molecule, which method comprises:
- (a) providing a sample of double stranded (ds) nucleic acid which contains the target sequence;
- (b) denaturing the sample so as to separate the two strands;
- (c) annealing to the sample a forward and a back oligonucleotide primer, the forward primer being specific for a sequence at or adjacent the 3′ end of the sense strand of the target sequence, the back primer being specific for a sequence at or adjacent the 3′ end of the antisense strand of the target sequence, under conditions which allow the primers to hybridize to the nucleic acid at or adjacent the target sequence;
- (d) treating the annealed sample with a DNA polymerase enzyme in the presence of deoxynucleoside triphosphates under conditions which cause primer extension to take place; and
- (e) denaturing the sample under conditions such that the extended primers become separated from the target sequence.
- Preferably, the method of the present invention further includes the step (f) of repeating steps (c) to (e) on the denatured mixture a plurality of times.
- Preferably, the method of the present invention is used to clone complete variable domains from Ig molecules, most preferably from Ig heavy chains. In the most preferred instance, the method will produce a DNA sequence encoding a ligand according to the present invention.
- In step (c) recited above, the forward primer becomes annealed to the sense strand of the target sequence at or adjacent the 3′ end of the strand. In a similar manner, the back primer becomes annealed to the antisense strand of the target sequence at or adjacent the 3′ end of the strand. Thus, the forward primer anneals at or adjacent the region of the ds nucleic acid which encodes the C-terminal end of the variable region or domain. Similarly, the back primer anneals at or adjacent the region of the ds nucleic acid which encodes the N-terminal end of the variable domain.
- In step (d), nucleotides are added onto the 3′ end of the forward and back primers in accordance with the sequence of the strand to which they are annealed. Primer extension will continue in this manner until stopped by the beginning of the denaturing step (e). It must therefore be ensured that step (d) is carried out for a long enough time to ensure that the primers are extended so that the extended strands totally overlap one another.
- In step (e), the extended primers are separated from the ds nucleic acid. The ds nucleic acid can then serve again as a substrate to which further primers can anneal. Moreover, the extended primers themselves have the necessary complementary sequences to enable the primers to anneal thereto.
- During further cycles, if step (f) is used, the amount of extended primers will increase exponentially so that at the end of the cycles there will be a large quantity of cDNA having sequences complementary to the sense and antisense strands of the target sequence. Thus, the method of the present invention will result in the accumulation of a large quantity of cDNA which can form ds cDNA encoding at least part of the variable domain.
- As will be apparent to the skilled person, some of the steps in the method may be carried out simultaneously or sequentially as desired.
- The forward and back primers may be provided as isolated oligonucleotides, in which case only two oligonucleotides will be used. However, alternatively the forward and back primers may each be supplied as a mixture of closely related oligonucleotides. For instance, it may be found that at a particular point in the sequence to which the primer is to anneal, there is the possibility of nucleotide variation. In this case a primer may be used for each possible nucleotide variation. Furthermore it may be possible to use two or more sets of “nested” primers in the method to enhance the specific cloning of variable region genes.
- The method described above is similar to the method described by Saiki et al. [17]. A similar method is also used in the methods described in EP-
A-0 200 362. In both cases the method described is carried out using primers which are known to anneal efficiently to the specified nucleotide sequence. In neither of these disclosures was it suggested that the method could be used to clone Ig parts of variable domain encoding sequences, where the target sequence contains inherently highly variable areas. - The ds nucleic acid sequence used in the method of the present invention may be derived from mRNA. For instance, RNA may be isolated in known manner from a cell or cell line which is known to produce Igs. mRNA may be separated from other RNA by oligo-dT chromatography. A complementary strand of cDNA may then be synthesized on the mRNA template, using reverse transcriptase and a suitable primer, to yield an RNA/DNA heteroduplex. A second strand of DNA can be made in one of several ways, for example, by priming with RNA fragments of the mRNA strand (made by incubating RNA/DNA heteroduplex with RNase H) and using DNA polymerase, or by priming with a synthetic oligodeoxynucleotide primer which anneals to the 3′ end of the first strand and using DNA polymerase. It has been found that the method of the present invention can be carried out using ds cDNA prepared in this way.
- When making such ds cDNA, it is possible to use a forward primer which anneals to a sequence in the CH1 domain (for a heavy chain variable domain) or the Cλ or Cκ domain (for a light chain variable domain). These will be located in close enough proximity to the target sequence to allow the sequence to be cloned.
- The back primer may be one which anneals to a sequence at the N-terminal end of the VH1, Vκ or V λ domain. The back primer may consist of a plurality of primers having a variety of sequences designed to be complementary to the various families of VH1, Vκ or Vλ sequences known. Alternatively the back primer may be a single primer having a consensus sequence derived from all the families of variable region genes.
- Surprisingly, it has been found that the method of the present invention can be carried out using genomic DNA. If genomic DNA is used, there is a very large amount of DNA present, including actual coding sequences, introns and untranslated sequences between genes. Thus, there is considerable scope for non-specific annealing under the conditions used. However, it has surprisingly been found that there is very little non-specific annealing. It is therefore unexpected that it has proved possible to clone the genes of Ig-variable domains from genomic DNA.
- Under some circumstances the use of genomic DNA may prove advantageous compared with use of mRNA, as the mRNA is readily degraded, and especially difficult to prepare from clinical samples of human tissue.
- Thus, in accordance with an aspect of the present invention, the ds nucleic acid used in step (a) is genomic DNA.
- When using genomic DNA as the ds nucleic acid source, it will not be possible to use as the forward primer an oligonucleotide having a sequence complementary to a sequence in a constant domain. This is because, in genomic DNA, the constant domain genes are generally separated from the variable domain genes by a considerable number of base pairs. Thus, the site of annealing would be too remote from the sequence to be cloned.
- It should be noted that the method of the present invention can be used to clone both rearranged and unrearranged variable domain sequences from genomic DNA. It is known that in germ line genomic DNA the three genes, encoding the VH, DH and JH respectively, are separated from one another by considerable numbers of base pairs. On maturation of the immune response, these genes are rearranged so that the VH, DH and JH genes are fused together to provide the gene encoding the whole variable domain (see
FIG. 1 ). By using a forward primer specific for a sequence at or adjacent the 3′ end of the sense strand of the genomic “unrearranged” VH gene, it is possible to clone the “unrearranged” VH gene alone, without also cloning the DH and JH genes. This can be of use in that it will then be possible to fuse the VH gene onto pre-cloned or synthetic DH and DH genes. In this way, rearrangement of the variable domain genes can be carried out in vitro. - The oligonucleotide primers used in step (c) may be specifically designed for use with a particular target sequence. In this case, it will be necessary to sequence at least the 5′ and 3′ ends of the target sequence so that the appropriate oligonucleotides can be synthesized. However, the present inventors have discovered that it is not necessary to use such specifically designed primers. Instead, it is possible to use a species specific general primer or a mixture of such primers for annealing to each end of the target sequence. This is not particularly surprising as regards the 3′ end of the target sequence. It is known that this end of the variable domain encoding sequence leads into a segment encoding JH which is known to be relatively conserved. However, it was surprisingly discovered that, within a single species, the sequence at the 5′ end of the target sequence is sufficiently well conserved to enable a species specific general primer or a mixture thereof to be designed for the 5′ end of the target sequence.
- Therefore according to a preferred aspect of the present invention, in step (c) the two primers which are used are species specific general primers, whether used as single primers or as mixtures of primers. This greatly facilitates the cloning of any undetermined target sequence since it will avoid the need to carry out any sequencing on the target sequence in order to produce target sequence-specific primers. Thus the method of this aspect of the invention provides a general method for cloning variable region or domain encoding sequences of a particular species.
- Once the variable domain gene has been cloned using the method described above, it may be directly inserted into an expression vector, for instance using the PCR reaction to paste the gene into a vector.
- Advantageously, however, each primer includes a sequence including a restriction enzyme recognition site. The sequence recognized by the restriction enzyme need not be in the part of the primer which anneals to the ds nucleic acid, but may be provided as an extension which does not anneal. The use of primers with restriction sites has the advantage that the DNA can be cut with at least one restriction enzyme which leaves 3′ or 5′ overhanging nucleotides. Such DNA is more readily cloned into the corresponding sites on the vectors than blunt end fragments taken directly from the method. The ds cDNA produced at the end of the cycles will thus be readily insertable into a cloning vector by use of the appropriate restriction enzymes. Preferably the choice of restriction sites is such that the ds cDNA is cloned directly into an expression vector, such that the ligand encoded by the gene is expressed. In this case the restriction site is preferably located in the sequence which is annealed to the ds nucleic acid.
- Since the primers may not have a sequence exactly complementary to the target sequence to which it is to be annealed, for instance because of nucleotide variations or because of the introduction of a restriction enzyme recognition site, it may be necessary to adjust the conditions in the annealing mixture to enable the primers to anneal to the ds nucleic acid. This is well within the competence of the person skilled in the art and needs no further explanation.
- In step (d), any DNA polymerase may be used. Such polymerases are known in the art and are available commercially. The conditions to be used with each polymerase are well known and require no further explanation here. The polymerase reaction will need to be carried out in the presence of the four nucleoside triphosphates. These and the polymerase enzyme may already be present in the sample or may be provided afresh for each cycle.
- The denaturing step (e) may be carried out, for instance, by heating the sample, by use of chaotropic agents, such as urea or guanidine, or by the use of changes in ionic strength or pH. Preferably, denaturing is carried out by heating since this is readily reversible. Where heating is used to carry out the denaturing, it will be usual to use a thermostable DNA polymerase, such as Taq polymerase, since this will not need replenishing at each cycle.
- If heating is used to control the method, a suitable cycle of heating comprises denaturation at about 95° C. for about 1 minute, annealing at from 30° C. to 65° C. for about 1 minute and primer extension at about 75° C. for about 2 minutes. To ensure that elongation and renaturation is complete, the mixture after the final cycle is preferably held at about 60° C. for about 5 minutes.
- The product ds cDNA may be separated from the mixture for instance by gel electrophoresis using agarose gels. However, if desired, the ds cDNA may be used in unpurified form and inserted directly into a suitable cloning or expression vector by conventional methods. This will be particularly easy to accomplish if the primers include restriction enzyme recognition sequences.
- The method of the present invention may be used to make variations in the sequences encoding the variable domains. For example this may be achieved by using a mixture of related oligonucleotide primers as at least one of the primers. Preferably the primers are particularly variable in the middle of the primer and relatively conserved at the 5′ and 3′ ends. Preferably the ends of the primers are complementary to the framework regions of the variable domain, and the variable region in the middle of the primer covers all or part of a CDR. Preferably a forward primer is used in the area which forms the third CDR. If the method is carried out using such a mixture of oligonucleotides, the product will be a mixture of variable domain encoding sequences. Moreover, variations in the sequence may be introduced by incorporating some mutagenic nucleotide triphosphates in step (d), such that point mutations are scattered throughout the target region. Alternatively such point mutations are introduced by performing a large number of cycles of amplification, as errors due to the natural error rate of the DNA polymerase are amplified, particularly when using high concentrations of nucleoside triphosphates.
- The method of this aspect of the present invention has the advantage that it greatly facilitates the cloning of variable domain encoding sequences directly from mRNA or genomic DNA. This in turn will facilitate the production of modified Ig-type molecules by any of the prior art methods referred to above. Further, target genes can be cloned from tissue samples containing antibody producing cells, and the genes can be sequenced. By doing this, it will be possible to look directly at the immune repertoire of a patient. This “fingerprinting” of a patient's immune repertoire could be of use in diagnosis, for instance of auto-immune diseases.
- In the method for amplifying the amount of a gene encoding a variable domain, a single set of primers is used in several cycles of copying via the polymerase chain reaction. As a less preferred alternative, there is provided a second method which comprises steps (a) to (d) as above, which further includes the steps of:
- (g) treating the sample of ds cDNA with traces of DNAse in the presence of DNA polymerase I to allow nick translation of the DNA; and
- (h) cloning the ds cDNA into a vector.
- If desired, the second method may further include the steps of:
- (i) digesting the DNA of recombinant plasmids to release DNA fragments containing genes encoding variable domains; and
- (j) treating the fragments in a further set of steps (c) to (h).
- Preferably the fragments are separated from the vector and from other fragments of the incorrect size by gel electrophoresis.
- The steps (a) to (d) then (g) to (h) can be followed once, but preferably the entire cycle (c) to (d) and (g) to (h) is repeated at least once. In this way a priming step, in which the genes are specifically copied, is followed by a cloning step, in which the amount of genes is increased.
- In step (a) the ds cDNA is derived from mRNA. For Ig derived variable domains, the mRNA is preferably be isolated from lymphocytes which have been stimulated to enhance production of mRNA.
- In each step (c) the set of primers are preferably different from the previous step (c), so as to enhance the specificity of copying. Thus the sets of primers form a nested set. For example, for cloning of Ig heavy chain variable domains, the first set of primers may be located within the signal sequence and constant region, as described by Larrick et al., [18], and the second set of primers entirely within the variable region, as described by Orlandi et al., [19]. Preferably the primers of step (c) include restriction sites to facilitate subsequent cloning. In the last cycle the set of primers used in step (c) should preferably include restriction sites for introduction into expression vectors. In step (g) possible mismatches between the primers and the template strands are corrected by “nick translation”. In step (h), the ds cDNA is preferably cleaved with restriction enzymes at sites introduced into the primers to facilitate the cloning.
- According to another aspect of the present invention the product ds cDNA is cloned directly into an expression vector. The host may be prokaryotic or eukaryotic, but is preferably bacterial. Preferably the choice of restriction sites in the primers and in the vector, and other features of the vector will allow the expression of complete ligands, while preserving all those features of the amino acid sequence which are typical of the (methoded) ligands. For example, for expression of the rearranged variable genes, the primers would be chosen to allow the cloning of target sequences including at least all the three CDR sequences. The cloning vector would then encode a signal sequence (for secretion of the ligand), and sequences encoding the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the last (fourth) framework region.
- For expression of unrearranged VH genes as part of complete ligands, the primers would be chosen to allow the cloning of target sequences including at least the first two CDRs. The cloning vector could then encode signal sequence, the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the third framework region, the third CDR and fourth framework region.
- Primers and cloning vectors may likewise be devised for expression of single CDRs, particularly the third CDR, as parts of complete ligands. The advantage of cloning repertoires of single CDRs would permit the design of a “universal” set of framework regions, incorporating desirable properties such as solubility.
- Single ligands could be expressed alone or in combination with a complementary variable domain. For example, a heavy chain variable domain can be expressed either as an individual domain or, if it is expressed with a complementary light chain variable domain, as an antigen binding site. Preferably the two partners would be expressed in the same cell, or secreted from the same cell, and the proteins allowed to associate non-covalently to form an Fv fragment. Thus the two genes encoding the complementary partners can be placed in tandem and expressed from a single vector, the vector including two sets of restriction sites. Preferably the genes are introduced sequentially: for example the heavy chain variable domain can be cloned first and then the light chain variable domain. Alternatively the two genes are introduced into the vector in a single step, for example by using the polymerase chain reaction to paste together each gene with any necessary intervening sequence, as essentially described by Yon and Fried [29]. The two partners could be also expressed as a linked protein to produce a single chain Fv fragment, using similar vectors to those described above. As a further alternative the two genes may be placed in two different vectors, for example in which one vector is a phage vector and the other is a plasmid vector.
- Moreover, the cloned ds cDNA may be inserted into an expression vector already containing sequences encoding one or more constant domains to allow the vector to express Ig-type chains. The expression of Fab fragments, for example, would have the advantage over Fv fragments that the heavy and light chains would tend to associate through the constant domains in addition to the variable domains. The final expression product may be any of the modified Ig-type molecules referred to above.
- The cloned sequence may also be inserted into an expression vector so that it can be expressed as a fusion protein. The variable domain encoding sequence may be linked directly or via a linker sequence to a DNA sequence encoding any protein effector molecule, such as a toxin, enzyme, label or another ligand. The variable domain sequences may also be linked to proteins on the outer side of bacteria or phage. Thus, the method of this aspect of the invention may be used to produce receptors according to the invention.
- According to another aspect of the invention, the cloning of ds cDNA directly for expression permits the rapid construction of expression libraries which can be screened for binding activities. For Ig heavy and light chain variable genes, the ds cDNA may comprise variable genes isolated as complete rearranged genes from the animal, or variable genes built from several different sources, for example a repertoire of unrearranged VH genes combined with a synthetic repertoire of DH and JH genes. Preferably repertoires of genes encoding Ig heavy chain variable domains are prepared from lymphocytes of animals immunized with an antigen.
- The screening method may take a range of formats well known in the art. For example Ig heavy chain variable domains secreted from bacteria may be screened by binding to antigen on a solid phase, and detecting the captured domains by antibodies. Thus the domains may be screened by growing the bacteria in liquid culture and binding to antigen coated on the surface of ELISA plates. However, preferably bacterial colonies (or phage plaques) which secrete ligands (or modified ligands, or ligand fusions with proteins) are screened for antigen binding on membranes. Either the ligands are bound directly to the membranes (and for example detected with labelled antigen), or captured on antigen coated membranes (and detected with reagents specific for ligands). The use of membranes offers great convenience in screening many clones, and such techniques are well known in the art.
- The screening method may also be greatly facilitated by making protein fusions with the ligands, for example by introducing a peptide tag which is recognized by an antibody at the N-terminal or C-terminal end of the ligand, or joining the ligand to an enzyme which catalyses the conversion of a colorless substrate to a colored product. In the latter case, the binding of antigen may be detected simply by adding substrate. Alternatively, for ligands expressed and folded correctly inside eukaryotic cells, joining of the ligand and a domain of a transcriptional activator such as the GAL4 protein of yeast, and joining of antigen to the other domain of the GAL4 protein, could form the basis for screening binding activities, as described by Fields and Song [21].
- The preparation of proteins, or even cells with multiple copies of the ligands, may improve the avidity of the ligand for immobilized antigen, and hence the sensitivity of the screening method. For example, the ligand may be joined to a protein subunit of a multimeric protein, to a phage coat protein or to an outer membrane protein of E. coli such as ompA or lamB. Such fusions to phage or bacterial proteins also offers possibilities of selecting bacteria displaying ligands with antigen binding activities. For example such bacteria may be precipitated with antigen bound to a solid support, or may be subjected to affinity chromatography, or may be bound to larger cells or particles which have been coated with antigen and sorted using a fluorescence activated cell sorter (FACS). The proteins or peptides fused to the ligands are preferably encoded by the vector, such that cloning of the ds cDNA repertoire creates the fusion product.
- In addition to screening for binding activities of single ligands, it may be necessary to screen for binding or catalytic activities of associated ligands, for example, the associated Ig heavy and light chain variable domains. For example, repertoires of heavy and light chain variable genes may be cloned such that two domains are expressed together. Only some of the pairs of domains may associate, and only some of these associated pairs may bind to antigen. The repertoires of heavy and light chain variable domains could be cloned such that each domain is paired at random. This approach may be most suitable for isolation of associated domains in which the presence of both partners is required to form a cleft. Alternatively, to allow the binding of hapten. Alternatively, since the repertoires of light chain sequences are less diverse than those of heavy chains, a small repertoire of light chain variable domains, for example including representative members of each family of domains, may be combined with a large repertoire of heavy chain variable domains.
- Preferably however, a repertoire of heavy chain variable domains is screened first for antigen binding in the absence of the light chain partner, and then only those heavy chain variable domains binding to antigen are combined with the repertoire of light chain variable domains. Binding of associated heavy and light chain variable domains may be distinguished readily from binding of single domains, for example by fusing each domain to a different C-terminal peptide tag which are specifically recognized by different monoclonal antibodies.
- The hierarchical approach of first cloning heavy chain variable domains with binding activities, then cloning matching light chain variable domains may be particularly appropriate for the construction of catalytic antibodies, as the heavy chain may be screened first for substrate binding. A light chain variable domain would then be identified which is capable of association with the heavy chain, and “catalytic” residues such as cysteine or histidine (or prosthetic groups) would be introduced into the CDRs to stabilize the transition state or attack the substrate, as described by Baldwin and Schultz [22].
- Although the binding activities of non-covalently associated heavy and light chain variable domains (Fv fragments) may be screened, suitable fusion proteins may drive the association of the variable domain partners. Thus Fab fragments are more likely to be associated than the Fv fragments, as the heavy chain variable domain is attached to a single heavy chain constant domain, and the light chain variable domain is attached to a single light chain variable domain, and the two constant domains associate together.
- Alternatively the heavy and light chain variable domains are covalently linked together with a peptide, as in the single chain antibodies, or peptide sequences attached, preferably at the C-terminal end which will associate through forming cysteine bonds or through non-covalent interactions, such as the introduction of “leucine zipper” motifs. However, in order to isolate pairs of tightly associated variable domains, the Fv fragments are preferably used.
- The construction of Fv fragments isolated from a repertoire of variable region genes offers a way of building complete antibodies, and an alternative to hybridoma technology. For example by attaching the variable domains to light or suitable heavy chain constant domains, as appropriate, and expressing the assembled genes in mammalian cells, complete antibodies may be made and should possess natural effector functions, such as complement lysis. This route is particularly attractive for the construction of human monoclonal antibodies, as hybridoma technology has proved difficult, and for example, although human peripheral blood lymphocytes can be immortalized with Epstein Barr virus, such hybridomas tend to secrete low affinity IgM antibodies.
- Moreover, it is known that immunological mechanisms ensure that lymphocytes do not generally secrete antibodies directed against host-proteins. However it is desirable to make human antibodies directed against human proteins, for example to human cell surface markers to treat cancers, or to histocompatibility antigens to treat auto-immune diseases. The construction of human antibodies built from the combinatorial repertoire of heavy and light chain variable domains may overcome this problem, as it will allow human antibodies to be built with specificities which would normally have been eliminated.
- The method also offers a new way of making bispecific antibodies. Antibodies with dual specificity can be made by fusing two hybridomas of different specificities, so as to make a hybrid antibody with an Fab arm of one specificity, and the other Fab arm of a second specificity. However the yields of the bispecific antibody are low, as heavy and light chains also find the wrong partners. The construction of Fv fragments which are tightly associated should preferentially drive the association of the correct pairs of heavy with light chains. (It would not assist in the correct pairing of the two heavy chains with each other.) The improved production of bispecific antibodies would have a variety of applications in diagnosis and therapy, as is well known.
- Thus the invention provides a species specific general oligonucleotide primer or a mixture of such primers useful for cloning variable domain encoding sequences from animals of that species. The method allows a single pair or pair of mixtures of species specific general primers to be used to clone any desired antibody specificity from that species. This eliminates the need to carry out any sequencing of the target sequence to be cloned and the need to design specific primers for each specificity to be recovered.
- Furthermore it provides for the construction of repertoires of variable genes, for the expression of the variable genes directly on cloning, for the screening of the encoded domains for binding activities and for the assembly of the domains with other variable domains derived from the repertoire.
- Thus the use of the method of the present invention will allow for the production of heavy chain variable domains with binding activities and variants of these domains. It allows for the production of monoclonal antibodies and bispecific antibodies, and will provide an alternative to hybridoma technology. For instance, mouse splenic ds mRNA or genomic DNA may be obtained from a hyper-immunized mouse. This could be cloned using the method of the present invention and then the cloned ds DNA inserted into a suitable expression vector. The expression vector would be used to transform a host cell, for instance a bacterial cell, to enable it to produce an Fv fragment or a Fab fragment. The Fv or Fab fragment would then be built into a monoclonal antibody by attaching constant domains and expressing it in mammalian cells.
- In the Examples described below, the following oligonucleotide primers, or mixed primers were used. Their locations are marked on
FIG. 1 and sequences are as follows: -
VH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCCAG 3′; VH1FOR-2 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCC 3′; Hu1VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu2VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu3VHFOR 5′ CTTGGTGGATGCTGAGGAGACGGTGACC 3′; Hu4VHFOR 5′ CTTGGTGGATGCTGATGAGACGGTGACC 3′; MOJH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTGCGCCCCAG 3′; MOJH2FOR 5′ TGAGGAGACGGTGACCGTGGTGCCTTGGCCCCAG 3′; MOJH3FOR 5′ TGCAGAGACGGTGACCAGAGTCCCTTGGCCCCAG 3′; MOJH4FOR 5′ TGAGGAGACGGTGACCGAGGTTCCTTGACCCCAG 3′; HUJH1FOR 5′ TGAGGAGACGGTGACCAGGGTGCCCTGGCCCCAG 3′; HUJH2FOR 5′ TGAGGAGACGGTGACCAGGGTGCCACGGCCCCAG 3′; HUJH4FOR 5′ TGAGGAGACGGTGACCAGGGTTCCTTGGCCCCAG 3′; VK1FOR 5′ GTTAGATCTCCAGCTTGGTCCC 3′; VK2FOR 5′ CGTTAGATCTCCAGCTTGGTCCC 3′; VK3FOR 5′ CCGTTTCAGCTCGAGCTTGGTCCC 3′; MOJK1FOR 5′ CGTTAGATCTCCAGCTTGGTGCC 3′; MOJK3FOR 5′ GGTTAGATCTCCAGTCTGGTCCC 3′; MOJK4FOR 5′ CGTTAGATCTCCAACTTTGTCCC 3′; HUJK1FOR 5′ CGTTAGATCTCCACCTTGGTCCC 3′; HUJK3FOR 5′ CGTTAGATCTCCACTTTGGTCCC 3′; HUJK4FOR 5′ CGTTAGATCTCCACCTTGGTCCC 3′; HUJK5FOR 5′ CGTTAGATCTCCAGTCGTGTCCC 3′; VH1BACK 5′ AGGT(C/G)(C/A)A(G/A)CTGCAG(G/C)AGTC(T/A)GG 3′; Hu2VHIBACK: 5′ CAGGTGCAGCTGCAGCAGTCTGG 3′; HuVHIIBACK: 5′ CAGGTGCAGCTGCAGGAGTCGGG 3′; Hu2VHIIIBACK: 5′ GAGGTGCAGCTGCAGGAGTCTGG 3′; HuVHIVBACK: 5′ CAGGTGCAGCTGCAGCAGTCTGG 3′; MOVHIBACK 5′ AGGTGCAGCTGCAGGAGTCAG 3′; MOVHIIABACK 5′ AGGTCCAGCTGCAGCA(G/A)TCTGG 3′; MOVHIIBBACK 5′ AGGTCCAACTGCAGCAGCCTGG 3′; MOVHIIBACK 5′ AGGTGAAGCTGCAGGAGTCTGG 3′; VK1BACK 5′ GACATTCAGCTGACCCAGTCTCCA 3′; VK2BACK 5′ GACATTGAGCTCACCCAGTCTCCA 3′; MOVKIIABACK 5′ GATGTTCAGCTGACCCAAACTCCA 3′ MOVKIIBBACK 5′ GATATTCAGCTGACCCAGGATGAA 3′; HuHep1FOR 5′ C(A/G)(C/G)TGAGCTCACTGTGTCTCTCGCACA 3′; HuOcta1BACK 5′ CGTGAATATGCAAATAA 3′; HUOcta2BACK 5′ AGTAGGAGACATGCAAAT 3′; and HuOcta3BACK 5′ CACCACCCACATGCAAAT 3′; VHMUT1 5′ GGAGACGGTGACCGTGGTCCCTTGGCCCCAGTAGTCAAGNNNNNNNN NNNNCTCTCTGGC 3′ (where N is an equimolar mixture of T, C, G and A) M13 pRIMER 5′ AACAGCTATGACCATG 3′ (New England Biolabs *1201) - VH1FOR is designed to anneal with the 3′ end of the sense strand of any mouse heavy chain variable domain encoding sequence. It contains a BstEII recognition site. VK1FOR is designed to anneal with the 3′ end of the sense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a BglII recognition site. VH1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse heavy chain variable domain and contains a PstI recognition site. VK1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a PvuII recognition site.
- In this Example five mouse hybridomas were used as a source of ds nucleic acid. The hybridomas produce monoclonal antibodies (MAbs) designated MBr1 [23], BW431/26 [24], BW494/32 [25], BW250/183 [24,26] and BW704/152 [27]. MAb MBr1 is particularly interesting in that it is known to be specific for a saccharide epitope on a human mammary carcinoma line MCF-7 [28].
- Cloning Via mRNA
- Each of the five hybridomas referred to above was grown up in roller bottles and about 5×108 cells of each hybridoma were used to isolate RNA. mRNA was separated from the isolated RNA using oligodT cellulose [29]. First strand cDNA was synthesized according to the procedure described by Maniatis et al. [30] as set out below.
- In order to clone the heavy chain variable domain encoding sequence, a 50 μl reaction solution which contains 10 μg mRNA, 20 pmole VH1FOR primer, 250 μM each of dATP, dTTP, dCTP and dGTP, 10 mM dithiothreitol (DTT), 100 mM Tris.HCl, 10 MM MgCl2 and 140 mM KCl, adjusted to pH 8.3 was prepared. The reaction solution was heated at 70° C. for ten minutes and allowed to cool to anneal the primer to the 3′ end of the variable domain encoding sequence in the mRNA. To the reaction solution was then added 46 units of reverse transcriptase (Anglian Biotec) and the solution was then incubated at 42° C. for 1 hour to cause first strand cDNA synthesis.
- In order to clone the light chain variable domain encoding sequence, the same procedure as set out above was used except that the VK1FOR primer was used in place of the VH1 FOR primer.
- Amplification from RNA/DNA Hybrid
- Once the ds RNA/DNA hybrids had been produced, the variable domain encoding sequences were amplified as follows. For heavy chain variable domain encoding sequence amplification, a 50 μl reaction solution containing 5 μl of the ds RNA/DNA hybrid-containing solution, 25 pmole each of VH1FOR and VH1BACK primers, 250 μM of dATP, dTTP, dCTP and dGTP, 67 mM Tris.HCl, 17 mM ammonium sulphate, 10 mM MgCl2, 200 μg/ml gelatine and 2 units Taq polymerase (Cetus) was prepared. The reaction solution was overlaid with paraffin oil and subjected to 25 rounds of temperature cycling using a Techne PHC-1 programmable heating block. Each cycle consisted of 1 minute and 95° C. (to denature the nucleic acids), 1 minute at 30° C. (to anneal the primers to the nucleic acids) and 2 minutes at 72° C. (to cause elongation from the primers). After the 25 cycles, the reaction solution and the oil were extracted twice with ether, once with phenol and once with phenol/CHCl3. Thereafter ds cDNA was precipitated with ethanol. The precipitated ds cDNA was then taken up in 50 μl of water and frozen.
- The procedure for light chain amplification was exactly as described above, except that the VK1FOR and VK1BACK primers were used in place of the VH1FOR and VH1BACK primers respectively.
- 5 μl of each sample of amplified cDNA was fractionated on 2% agarose gels by electrophoresis and stained with ethidium bromide. This showed that the amplified ds cDNA gave a major band of the expected size (about 330 bp). (However the band for VK 0 DNA of MBr1 was very weak. It was therefore excised from the gel and reamplified in a second round.) Thus by this simple procedure, reasonable quantities of ds DNA encoding the light and heavy chain variable domains of the five MAbs were produced.
- Heavy Chain Vector Construction
- A BstEII recognition site was introduced into the vector M13-HuVHNP [31] by site directed mutagenesis [32,33] to produce the vector M13-VHPCR1 (
FIGS. 2 and 3 ). - Each amplified heavy chain variable domain encoding sequence was digested with the restriction enzymes PstI and BstEII. The fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VHPCR1 which had been digested with PstI and BstEII and purified on an 0.8% agarose gel. Clones containing the variable domain inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding variable gene in the M13-VHPCR1 vector.
- There is an internal PstI site in the heavy chain variable domain encoding sequences of BW431/26. This variable domain encoding sequence was therefore assembled in two steps. The 3′ PstI-BstEII fragment was first cloned into M13-VHPCR1, followed in a second step by the 5′ PstI fragment.
- Light Chain Vector Construction
- Vector M13 mp 18 [35] was cut with PvuII and the vector backbone was blunt ligated to a synthetic HindIII-BamHI polylinker. Vector M13-HuVKLYS [36] was digested with HindIII and BamHI to isolate the HuVKLYS gene. This HindIII-BamHI fragment was then inserted into the HindIII-BamHI polylinker site to form a vector M13-VKPCR1 which lacks any PvuII sites in the vector backbone (
FIGS. 4 and 5 ). This vector was prepared in E. coli JM110 [22] to avoid dam methylation at the BclI site. - Each amplified light chain variable domain encoding sequence was digested with PvuII and BglII. The fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VKPCR1 which had been digested with PvuII and BclI, purified on an 0.8% agarose gel and treated with calf intestinal phosphatase. Clones containing the light chain variable region inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding region of the variable domain in the M13-VKPCR1 vector.
- The nucleotide sequences of the MBr1 heavy and light chain variable domains are shown in
FIG. 6 with part of the flanking regions of the M13-VHPCR1 and M13-VKPCR1 vectors. - Antibody Expression
- The HindIII-BamHI fragment carrying the MBr1 heavy chain variable domain encoding sequence in M13-VHPCR1 was recloned into a pSV-gpt vector with human γ1 constant regions [37] (
FIG. 7 ). The MBr1 light chain variable domain encoding sequence in M13-VKPCR1 was recloned as a HindIII-BamHI fragment into a pSV vector, PSV-hyg-HuCK with a hygromycin resistance marker and a human kappa constant domain (FIG. 8 ). The assembly of the genes is summarized inFIG. 9 . - The vectors thus produced were linearized with PvuI (in the case of the pSV-hygro vectors the PvuI digest is only partial) and cotransfected into the non-secreting mouse myeloma line NSO [38] by electroporation [39]. One day after cotransfection, cells were selected in 0.3 μg/ml mycophenolic acid (MPA) and after seven days in 1 μg/ml MPA. After 14 days, four wells, each containing one or two major colonies, were screened by incorporation of 14C-lysine [40] and the secreted antibody detected after precipitation with protein-A Sepharose™ (Pharmacia) on SDS-PAGE [41]. The gels were stained, fixed, soaked in a fluorographic reagent, Amplify™ (Amersham), dried and autoradiographed on preflashed film at −70° C. for 2 days.
- Supernatant was also tested for binding to the mammary carcinoma line MCF-7 and the colon carcinoma line HT-29, essentially as described by Menard et al. [23], either by an indirect immunofluorescence assay on cell suspensions (using a fluorescein-labelled goat anti-human IgG (Amersham)) or by a solid phase RIA on monolayers of fixed cells (using 125I-protein A (Amersham)).
- It was found that one of the supernatants from the four wells contained secreted antibody. The chimeric antibody in the supernatant, like the parent mouse MBr1 antibody, was found to bind to MCF-7 cells but not the HT-29 cells, thus showing that the specificity had been properly cloned and expressed.
- Preparation of DNA from Spleen
- The DNA from the mouse spleen was prepared in one of two ways (although other ways can be used).
-
Method 1. A mouse spleen was cut into two pieces and each piece was put into a standard Eppendorf tube with 200 μl of PBS. The tip of a 1 ml glass pipette was closed and rounded in the blue flame of a Bunsen burner. The pipette was used to squash the spleen piece in each tube. The cells thus produced were transferred to a fresh Eppendorf tube and the method was repeated three times until the connective tissue of the spleen appeared white. Any connective tissue which has been transferred with the cells was removed using a drawn-out Pasteur pipette. The cells were then washed in PBS and distributed into four tubes. - The mouse spleen cells were then sedimented by a 2 minute spin in a Microcentaur centrifuge at low speed setting. All the supernatant was aspirated with a drawn out Pasteur pipette. If desired, at this point the cell sample can be frozen and stored at −20° C.
- To the cell sample (once thawed if it had been frozen) was added 500 μl of water and 5 μl of a 10% solution of NP-40, a non-ionic detergent. The tube was closed and a hole was punched in the lid. The tube was placed on a boiling water bath for 5 minutes to disrupt the cells and was then cooled on ice for 5 minutes. The tube was then spun for 2 minutes at high speed to remove cell debris.
- The supernatant was transferred to a new tube and to this was added 125 μl 5M NaCl and 30 μl 1M MOPS adjusted to pH 7.0. The DNA in the supernatant was absorbed on a
Quiagen 5 tip and purified following the manufacturer's instructions for lambda DNA. After isopropanol precipitation, the DNA was resuspended in 500 μl water. -
Method 2. This method is based on the technique described in Maniatis et al. [30]. A mouse spleen was cut into very fine pieces and put into a 2 ml glass homogenizer. The cells were then freed from the tissue by several slow up and down strokes with the piston. The cell suspension was made in 500 μl phosphate buffered saline (PBS) and transferred to an Eppendorf tube. The cells were then spun for 2 min at low speed in a Microcentaur centrifuge. This results in a visible separation of white and red cells. The white cells, sedimenting slower, form a layer on top of the red cells. The supernatant was carefully removed and spun to ensure that all the white cells had sedimented. The layer of white cells was resuspended in two portions of 500 μl PBS and transferred to another tube. - The white cells were precipitated by spinning in the Microcentaur centrifuge at low speed for one minute. The cells were washed a further two times with 500 μl PBS, and were finally resuspended in 200 μl PBS. The white cells were added to 2.5
ml 25 mM EDTA and 10 mM Tris.Cl, pH 7.4, and vortexed slowly. Whilevortexing 25μl 20% SDS was added. The cells lysed immediately and the solution became viscous and clear. 100 μl of 20 mg/ml proteinase K was added and incubated one to three hours at 50° C. - The sample was extracted with an equal volume of phenol and the same volume of chloroform, and vortexed. After centrifuging, the aqueous phase was removed and 1/10 volume 3M ammonium acetate was added. This was overlaid with three volumes of cold ethanol and the tube rocked carefully until the DNA strands became visible. The DNA was spooled out with a Pasteur pipette, the ethanol allowed to drip off, and the DNA transferred to 1 ml of 10 mM Tris.Cl pH 7.4, 0.1 mM EDTA in an Eppendorf tube. The DNA was allowed to dissolve in the cold overnight on a roller.
- Amplification From Genomic DNA
- The DNA solution was diluted 1/10 in water and boiled for 5 min prior to using the polymerase chain reaction (PCR). For each PCR reaction, typically 50-200 ng of DNA were used.
- The heavy and light chain variable domain encoding sequences in the genomic DNA isolated from the human PBL or the mouse spleen cells was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA Hybrid” in Example 1, except that during the annealing part of each cycle, the temperature was held at 65° C. and that 30 cycles were used. Furthermore, to minimize the annealing between the 3′ ends of the two primers, the sample was first heated to 95° C., then annealed at 65° C., and only then was the Taq polymerase added. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- The primers used to amplify the mouse spleen genomic DNA were VH1FOR and VH1BACK, for the heavy chain variable domain and VK2FOR and VK1BACK, for the light chain variable domain. (VK2FOR only differs from VK1FOR in that it has an extra C residue on the 5′ end.)
- Other sets of primers, designed to optimize annealing with different families of mouse VH and Vκ genes were devised and used in mixtures with the primers above. For example, mixtures of VK1FOR, MOJK1FOR, MOJK3FOR and MOJK4FOR were used as forward primers and mixtures of VK1BACK, MOVKIIABACK and MOVKIIBBACK as back primers for amplification of Vκ genes. Likewise mixtures of VH1FOR, MOJH1FOR, MOJH2FOR, MOJH3FOR and MOJH4FOR were used as forward primers and mixtures of VH1BACK, MOVHIBACK, MOVHIIABACK, MOVHIIBBACK, MOVHIIIBACK were used as backward primers for amplification of VH genes.
- All these heavy chain FOR primers referred to above contain a BstEII site and all the BACK primers referred to above contain a PstI site. These light chain FOR and BACK primers referred to above all contain BglII and PvuII sites respectively. Light chain primers (VK3FOR and VK2BACK) were also devised which utilized different restriction sites, SacI and XhoI.
- Typically all these primers yielded amplified DNA of the correct size on gel electrophoresis, although other bands were also present. However, a problem was identified in which the 5′ and 3′ ends of the forward and backward primers for the VH genes were partially complementary, and this could yield a major band of “primer-dimer” in which the two oligonucleotides prime on each other. For this reason an improved forward primer, VH1FOR-2 was devised in which the two 3′ nucleotides were removed from VH1FOR.
- Thus, the preferred amplification conditions for mouse VH genes are as follows: the sample was made in a volume of 50-100 μl, 50-100 ng of DNA, VH1FOR-2 and VH1BACK primers (25 pmole of each), 250 μM of each deoxynucleotide triphosphate, 10 mM Tris.HCl, pH 8.8, 50 mM KCl, 1.5 mM MgCl2, and 100 μg/ml gelatine. The sample was overlaid with paraffin oil, heated to 95° C. for 2 min, 65° C. for 2 min, and then to 72° C.: taq polymerase was added after the sample had reached the elongation temperature and the reaction continued for 2 min at 72° C. The sample was subjected to a further 29 rounds of temperature cycling using the Techne PHC-1 programmable heating block.
- The preferred amplification conditions for mouse Vκ genes from genomic DNA are as follows: the sample treated as above except with Vκ primers, for example VK3FOR and VK2BACK, and using a cycle of 94° C. for one minute, 60° C. for one minute and 72° C. for one minute.
- The conditions which were devised for genomic DNA are also suitable for amplification from the cDNA derived from mRNA from mouse spleen or mouse hybridoma.
- Cloning and Analysis of Variable Region Genes
- The reaction mixture was then extracted twice with 40 μl of water-saturated diethyl ether. This was followed by a standard phenol extraction and ethanol precipitation as described in Example 1. The DNA pellet was then dissolved in 100
μl 10 mM Tris.Cl, 0.1 mM EDTA. - Each reaction mixture containing a light chain variable domain encoding sequence was digested with SacI and XhoI (or with PvuII and BglII) to enable it to be ligated into a suitable expression vector. Each reaction mixture containing a heavy chain variable domain encoding sequence was digested with PstI and BstEII for the same purpose.
- The heavy chain variable genes isolated as above from a mouse hyper-immunized with lysozyme were cloned into M13VHPCR1 vector and sequenced. The complete sequences of 48 VH gene clones were determined (
FIGS. 10 a-10 b). All but two of the mouse VH gene families were represented, with frequencies of: VA (1), IIIC (1), IIIB (8), IIIA (3), IIB (17), IIA (2), IB (12), IA (4). In 30 clones, the D segments could be assigned to families SP2 (14), FL16 (11) and Q52 (5), and in 38 clones the JH minigenes to families JH1 (3), JH2 (7), JH3 (14) and JH4 (14). The different sequences of CDR3 marked out each of the 48 clones as unique. Nine pseudogenes and 16 unproductive rearrangements were identified. Of the clones sequenced, 27 have open reading frames. - Thus the method is capable of generating a diverse repertoire of heavy chain variable genes from mouse spleen DNA.
- Preparation of mRNA
- Human peripheral blood lymphocytes were purified and mRNA prepared directly (Method 1), or mRNA was prepared after addition of Epstein Barr virus (Method 2).
-
Method 1. 20 ml of heparinized human blood from a healthy volunteer was diluted with an equal volume of phosphate buffered saline (PBS) and distributed equally into 50 ml Falcon tubes. The blood was then underlayed with 15 ml Ficoll Hypaque (Pharmacia 10-A-001-07). To separate the lymphocytes from the red blood cells, the tubes were spun for 10 minutes at 1800 rpm at room temperature in an IEC Centra 3E table centrifuge. The peripheral blood lymphocytes (PBL) were then collected from the interphase by aspiration with a Pasteur pipette. The cells were diluted with an equal volume of PBS and spun again at 1500 rpm for 15 minutes. The supernatant was aspirated, the cell pellet was resuspended in 1 ml PBS and the cells were distributed into two Eppendorf tubes. -
Method 2. 40 ml human blood from a patient with HIV in the pre-AIDS condition was layered on Ficoll to separate the white cells (seeMethod 1 above). The white cells were then incubated in tissue culture medium for 4-5 days. Onday 3, they were infected with Epstein Barr virus. The cells were pelleted (approx 2×107 cells) and washed in PBS. - The cells were pelleted again and lysed with 7 ml 5M guanidine isothiocyanate, 50 mM Tris, 10 mM EDTA, 0.1 mM dithiothreitol. The cells were vortexed vigorously and 7 volumes of 4M LiCl added. The mixture was incubated at 4° C. for 15-20 hrs. The suspension was spun and the supernatant resuspended in 3M LiCl and centrifuged again. The pellet was dissolved in 2 ml 0.1% SDS, 10 mM Tris HCl and 1 mM EDTA. The suspension was frozen at −20° C., and thawed by vortexing for 20 s every 10 min for 45 min. A large white pellet was left behind and the clear supernatant was extracted with phenol chloroform, then with chloroform. The RNA was precipitated by adding 1/10 volume 3M sodium acetate and 2 vol ethanol and leaving overnight at −20° C. The pellet was suspended in 0.2 ml water and reprecipitated with ethanol. Aliquots for cDNA synthesis were taken from the ethanol precipitate which had been vortexed to create a fine suspension.
- 100 μl of the suspension was precipitated and dissolved in 20 μl water for cDNA synthesis [30] using 10 pmole of a HUFOR primer (see below) in final volume of 50 μl. A sample of 5 μl of the cDNA was amplified as in Example 2 except using the primers for the human VH gene families (see below) using a cycle of 95° C., 60° C. and 72° C.
- The back primers for the amplification of human DNA were designed to match the available human heavy and light chain sequences, in which the different families have slightly different nucleotide sequences at the 5′ end. Thus for the human VH genes, the primers Hu2VHIBACK, HuVHIIBACK, Hu2VHIIIBACK and HuVH1VBACK were designed as back primers, and HUJH1FOR, HUJH2FOR and HUJH4FOR as forward primers based entirely in the variable gene. Another set of forward primers Hu1VHFOR, Hu2VHFOR, Hu3VHFOR, and Hu4VHFOR was also used, which were designed to match the human J-regions and the 5′ end of the constant regions of different human isotypes.
- Using sets of these primers it was possible to demonstrate a band of amplified ds cDNA by gel electrophoresis.
- One such experiment was analyzed in detail to establish whether there was a diverse repertoire in a patient with HIV infection. It is known that during the course of AIDS, that T-cells and also antibodies are greatly diminished in the blood. Presumably the repertoire of lymphocytes is also diminished. In this experiment, for the forward priming, an equimolar mixture of primers Hu1VHFOR, Hu2VHFOR, Hu3VHFOR, and Hu4VHFOR (in
PCR 25 pmole ofprimer 5′ ends) was used. For the back priming, the primers Hu2VHIBACK, HuVHIIBACK, Hu2VHIIIBACK and HuVH1VBACK were used separately in four separate primings. The amplified DNA from the separate primings was then pooled, digested with restriction enzymes PstI and BstEII as above, and then cloned into the vector M13VHPCR1 for sequencing. The sequences reveal a diverse repertoire (FIG. 11 ) at this stage of the disease. - For human Vκ genes the primers HuJK1FOR, HUJK3FOR; HUJK4FOR and HUJK5FOR were used as forward primers and VK1BACK as back primer. Using these primers it was possible to see a band of amplified ds cDNA of the correct size by gel electrophoresis.
- Human peripheral blood lymphocytes of a patient with non-Hodgkins lymphoma were prepared as in Example 3 (Method 1). The genomic DNA was prepared from the PBL using the technique described in Example 2 (Method 2). The VH region in the isolated genomic DNA was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA hybrid” in Example 1 above, except that during the annealing part of each cycle, the temperature was held at 55° C. and that 30 cycles were used. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- The forward primer used was HuHep1FOR, which contains a SacI site. This primer is designed to anneal to the 3′ end of the unrearranged human VH region gene, and in particular includes a sequence complementary to the last three codons in the VH region gene and nine nucleotides downstream of these three codons.
- As the back primer, an equimolar mixture of HuOcta1BACK, HuOcta2BACK and HuOcta3BACK was used. These primers anneal to a sequence in the promoter region of the genomic DNA VH gene (see
FIG. 1 ). 5 μl of the amplified DNA was checked on 2% agarose gels in TBE buffer and stained with ethidium bromide. A double band was seen of about 620 nucleotides which corresponds to the size expected for the unrearranged VH gene. The ds cDNA was digested with SacI and cloned into an M13 vector for sequencing. Although there are some sequences which are identical, a range of different unrearranged human VH genes were identified (FIG. 12 ). - The heavy chain variable domain (VHLYS) of the D1.3 (anti-lysozyme) antibody was cloned into a vector similar to that described previously [42] but under the control of the lac z promoter, such that the VHLYS domain is attached to a pelB leader sequence for export into the periplasm. The vector was constructed by synthesis of the pelB leader sequence [43], using overlapping oligonucleotides, and cloning into a pUC 19 vector [35]. The VHLYS domain of the D1.3 antibody was derived from a cDNA clone [44] and the construct (PSW1) sequenced (
FIG. 13 ). - To express both heavy and light chain variable domains together, the light chain variable region (VKLYS) of the D1.3 antibody was introduced into the pSW1 vector, with a pelB signal sequence to give the construct pSW2 (
FIGS. 14 a-14 b). - A strain of E. coli (BMH71-18) [45] was then transformed [46,47] with the plasmid pSW1 or pSW2, and colonies resistant to ampicillin (100 μg/ml) were selected on a rich (2×TY=per litre of water, 16 g Bacto-tryptone, 10 g yeast extract, 5 g NaCl) plate which contained 1% glucose to repress the expression of variable domain(s) by catabolite repression.
- The colonies were inoculated into 50
ml 2×TY (with 1% glucose and 100 μg/ml ampicillin) and grown in flasks at 37° C. with shaking for 12-16 hr. The cells were centrifuged, the pellet washed twice with 50 mM sodium chloride, resuspended in 2×TY medium containing 100 μg/ml ampicillin and the inducer IPTG (1 mM) and grown for a further 30 hrs at 37° C. The cells were centrifuged and the supernatant was passed through a Nalgene filter (0.45 μm) and then down a 1-5 ml lysozyme-Sepharose® affinity column (Pharmacia Fine Chemicals, Inc.). (The column was derived by coupling lysozyme at 10 mg/ml to CNBr activated Sepharose.) The column was first washed with phosphate buffered saline (PBS), then with 50 mM diethylamine to elute the VHLYS domain (from pSW1) or VHLYS in association with VKLYS (from pSW2). - The VHLYS and VKLYS domains were identified by SDS polyacrylamide electrophoresis as the correct size. In addition, N-terminal sequence determination of VHLYS and VKLYS isolated from a polyacrylamide gel showed that the signal peptide had been produced correctly. Thus both the Fv fragment and the VHLYS domains are able to bind to the lysozyme affinity column, suggesting that both retain at least some of the affinity of the original antibody.
- The size of the VHLYS domain was compared by FPLC with that of the Fv fragment on Superose 12. This indicates that the VHLYS domain is a monomer. The binding of the VHLYS and Fv fragment to lysozyme was checked by ELISA, and equilibrium and rapid reaction studies were carried out using fluorescence quench.
- The ELISA for lysozyme binding was undertaken as follows:
- (1) The plates (Dynatech Immulon) were coated with 200 μl per well of 300 μg/ml lysozyme in 50 mM NaHCO3, pH 9.6 overnight at room temperature;
- (2) The wells were rinsed with three washes of PBS, and blocked with 300 μl per well of 1% Sainsbury's instant dried skimmed milk powder in PBS for 2 hours at 37° C.;
- (3) The wells were rinsed with three washes of PBS and 200 μl of VHLYS or Fv fragment (VHLYS associated with VKLYS) were added and incubated for 2 hours at room temperature;
- (4) The wells were washed three times with 0.05
% Tween 20 in PBS and then three times with PBS to remove detergent; - (5) 200 μl of a suitable dilution (1:1000) of rabbit polyclonal antisera raised against the Fv fragment in 2% skimmed milk powder in PBS was added to each well and incubated at room temperature for 2 hours;
- (6) Washes were repeated as in (4);
- (7) 200 μl of a suitable dilution (1:1000) of goat anti-rabbit antibody (ICN Immunochemicals) coupled to horse radish peroxidase, in 2% skimmed milk powder in PBS, was added to each well and incubated at room temperature for 1 hour;
- (8) Washes were repeated as in (4); and
- (9) 200
ml μl 20% hydrogen peroxide: water per 10 ml) was added to each well and the color allowed to develop for up to 10 minutes at room temperature. - The reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid pH 4.3. ELISA plates were read in a Titertek Multiscan plate reader. Supernatant from the induced bacterial cultures of both pSW1 (VHLYS domain) or pSW2 (Fv fragment) was found to bind to lysozyme in the ELISA.
- The purified VHLYS and Fv fragments were titrated with lysozyme using fluorescence quench (Perkin Elmer LS5B Luminescence Spectrometer) to measure the stoichiometry of binding and the affinity constant for lysozyme [48,49]. The titration of the Fv fragment at a concentration of 30 nM indicates a dissociation constant of 2.8 nM using a Scatchard analysis.
- A similar analysis using fluorescence quench and a Scatchard plot was carried out for VHLYS, at a VHLYS concentration of 100 nM. The stoichiometry of antigen binding is about 1 mole of lysozyme per mole of VHLYS (calculated from plot). (The concentration of VH domains was calculated from optical density at 280 nM using the typical extinction coefficient for complete immunoglobulins.) Due to possible errors in measuring low optical densities and the assumption about the extinction coefficient, the stoichiometry was also measured more carefully. VHLYS was titrated with lysozyme as above using fluorescence quench. To determine the concentration of VHLYS a sample of the stock solution was removed, a known amount of norleucine added, and the sample subjected to quantitative amino acid analysis. This showed a stoichiometry of 1.2 mole of lysozyme per mole of VHLYS domain. The dissociation constant was calculated as about 12 nM.
- The on-rates for VHLYS and Fv fragments with lysozyme were determined by stopped-flow analysis (HI Tech Stop Flow SHU machine) under pseudo-first order conditions with the fragment at a ten fold higher concentration than lysozyme [50]. The concentration of lysozyme binding sites was first measured by titration with lysozyme using fluorescence quench as above. The on rates were calculated per mole of binding site (rather than amount of VHLYS protein). The on-rate for the Fv fragment was found to be 2.2×106 M−1 s−1 at 25° C. The on-rate for the VHLYS fragment found to be 3.8×106 M−1 s−1 and the off-rate 0.075 s−1 at 20° C. The calculated affinity constant is 19 nM. Thus the VHLYS binds to lysozyme with a dissociation constant of about 19 nM, compared with that of the Fv of 3 nM.
- A mouse was immunized with hen egg white lysozyme (100 μg i.p.
day 1 in complete Freunds adjuvant), after 14 days immunized i.p. again with 100 μg lysozyme with incomplete Freunds adjuvant, and onday 35 i.v. with 50 μg lysozyme in saline. On day 39, spleen was harvested. A second mouse was immunized with keyhole limpet hemocyanin (KLH) in a similar way. The DNA was prepared from the spleen according to Example 2 (Method 2). The VH genes were amplified according to the preferred method in Example 2. - Human peripheral blood lymphocytes from a patient infected with HIV were prepared as in Example 3 (Method 2) and mRNA prepared. The VH genes were amplified according to the method described in Example 3, using primers designed for human VH gene families.
- After the PCR, the reaction mixture and oil were extracted twice with ether, once with phenol and once with phenol/CHCl3. The double stranded DNA was then taken up in 50 μl of water and frozen. 5 μl was digested with PstI and BstEII (encoded within the amplification primers) and loaded on an agarose gel for electrophoresis. The band of amplified DNA at about 350 bp was extracted.
- Expression of Anti-Lysozyme Activities
- The repertoire of amplified heavy chain variable domains (from mouse immunized with lysozyme and from human PBLs) was then cloned directly into the expression vector pSW1HPOLYMYC. This vector is derived from pSW1 except that the VHLYS gene has been removed and replaced by a polylinker restriction site. A sequence encoding a peptide tag was inserted (
FIG. 15 ). Colonies were toothpicked into 1 ml cultures. After induction (see Example 5 for details), 10 μl of the supernatant from fourteen 1 ml cultures was loaded on SDS-PAGE gels and the proteins transferred electrophoretically to nitrocellulose. The blot was probed with antibody 9E10 directed against the peptide tag. - The probing was undertaken as follows. The nitrocellulose filter was incubated in 3% bovine serum albumin (BSA)/TBS buffer for 20 min (10×TBS buffer is 100 mM Tris.HCl, pH 7.4, 9% w/v NaCl). The filter was incubated in a suitable dilution of antibody 9E10 (about 1/500) in 3% BSA/TBS for 1-4 hrs. After three washes in TBS (100 ml per wash, each wash for 10 min), the filter was incubated with 1:500 dilution of anti-mouse antibody (peroxidase conjugated anti-mouse Ig (Dakopats)) in 3% BSA/TBS for 1-2 hrs. After three washes in TBS and 0.1% Triton X-100 (about 100 ml per wash, each wash for 10 min), a solution containing 10 ml chloronaphthol in methanol (3 mg/ml), 40 ml TBS and 50 μl hydrogen peroxide solution was added over the blot and allowed to react for up to 10 min. The substrate was washed out with excess water. The blot revealed bands similar in mobility to VHLYSMYC on the Western blot, showing that other VH domains could be expressed.
- Colonies were then toothpicked individually into wells of an ELISA plate (200 μl) for growth and induction. They were assayed for lysozyme binding with the 9E10 antibody (as in Examples 5 and 7). Wells with lysozyme-binding activity were identified. Two positive wells (of 200) were identified from the amplified mouse spleen DNA and one well from the human cDNA. The heavy chain variable domains were purified on a column of lysozyme-Sepharose. The affinity for lysozyme of the clones was estimated by fluorescence quench titration as >50 nM. The affinities of the two clones (VH3 and VH8) derived from the mouse genes were also estimated by stop flow analysis (ratio of koff/kon) as 12 nM and 27 nM respectively. Thus both these clones have a comparable affinity to the VHLYS domain. The encoded amino acid sequences of VH3 and VH8 are given in
FIG. 16 , and that of the human variable domain inFIG. 17 . - A library of VH domains made from the mouse immunized with lysozyme was screened for both lysozyme and keyhole limpet hemocyanin (KLH) binding activities. Two thousand colonies were toothpicked in groups of five into wells of ELISA plates, and the supernatants tested for binding to lysozyme coated plates and separately to KLH coated plates. Twenty one supernatants were shown to have lysozyme binding activities and two to have KLH binding activities. A second expression library, prepared from a mouse immunized with KLH was screened as above. Fourteen supernatants had KLH binding activities and a single supernatant had lysozyme binding activity.
- This shows that antigen binding activities can be prepared from single VH domains, and that immunization facilitates the isolation of these domains.
- Taking a single rearranged VH gene, it may be possible to derive entirely new antigen binding activities by extensively mutating each of the CDRs. The mutagenesis might be entirely random, or be derived from pre-existing repertoires of CDRs. Thus a repertoire of CDR3s might be prepared as in the preceding examples by using “universal” primers based in the flanking sequences, and likewise repertoires of the other CDRs (singly or in combination). The CDR repertoires could be stitched into place in the flanking framework regions by a variety of recombinant DNA techniques.
- CDR3 appears to be the most promising region for mutagenesis as CDR3 is more variable in size and sequence than
CDRs - Multiple mutations were made in CDR3. The polymerase chain reaction (PCR) and a highly degenerate primer were used to make the mutations and by this means the original sequence of CDR3 was destroyed. (It would also have been possible to construct the mutations in CDR3 by cloning a mixed oligonucleotide duplex into restriction sites flanking the CDR or by other methods of site-directed mutagenesis). Mutants expressing heavy chain variable domains with affinities for lysozyme were screened and those with improved affinities or new specificities were identified.
- The source of the heavy chain variable domain was an M113 vector containing the VHLYS gene. The body of the sequence encoding the variable region was amplified using the polymerase chain reaction (PCR) with the mutagenic primer VHMUT1 based in CDR3 and the M13 primer which is based in the M13 vector backbone. The mutagenic primer hypermutates the central four residues of CDR3 (Arg-Asp-Tyr-Arg). The PCR was carried out for 25 cycles on a Techne PHC-1 programmable heat block using 100 ng single stranded M13 mp19SWO template, with 25 pmol of VHMUT1 and the M13 primer, 0.5 mM each dNTP, 67 mM Tris.HCl, pH 8.8, 10 mM MgCl2, 17 mM (NH4)2SO4, 200 μg/ml gelatine and 2.5 units Taq polymerase in a final volume of 50 μl. The temperature regime was 95° C. for 1.5 min, 25° C. for 1.5 min and 72° C. for 3 min (However a range of PCR conditions could be used). The reaction products were extracted with phenol/chloroform, precipitated with ethanol and resuspended in 10 mM Tris. HCl and 0.1 mM EDTA, pH 8.0.
- The products from the PCR were digested with PstI and BstEII and purified on a 1.5% LGT agarose gel in Tris acetate buffer using Geneclean® (Bio 101, LaJolla). The gel purified band was ligated into pSW2HPOLY (
FIG. 19 ). (This vector is related to pSW2 except that the body of the VHLYS gene has been replaced by a polylinker.) The vector was first digested with BstEII and PstI and treated with calf-intestinal phosphatase. Aliquots of the reaction mix were used to transform E. coli BMH 71-18 to ampicillin resistance. Colonies were selected on ampicillin (100 μg/ml) rich plates containing glucose at 0.8% w/v. - Colonies resulting from transfection were picked in pools of five into two 96 well Corning microtitre plates, containing 200
μl 2×TY medium and 100 μl TY medium, 100 μg/ml ampicillin and 1% glucose. The colonies were grown for 24 hours at 37° C. and then cells were washed twice in 200μl 50 mM NaCl, pelleting the cells in an IEC Centra-3 bench top centrifuge with microtitre plate head fitting. Plates were spun at 2,500 rpm for 10 min at room temperature. Cells were resuspended in 200μl 2×TY, 100 μg/ml ampicillin and 1 mM IPTG (Sigma) to induce expression, and grown for a further 24 hr. - Cells were spun down and the supernatants used in ELISA with lysozyme coated plates and anti-idiotypic sera (raised in rabbits against the Fv fragment of the D1.3 antibody). Bound anti-idiotypic serum was detected using horse radish peroxidase conjugated to anti-rabbit sera (ICN Immunochemicals). Seven of the wells gave a positive result in the ELISA. These pools were restreaked for single colonies which were picked, grown up, induced in microtitre plates and rescreened in the ELISA as above. Positive clones were grown up at the 50 ml scale and expression was induced. Culture supernatants were purified as in Example 5 on columns of lysozyme-Sepharose and eluates analysed on SDS-PAGE and staining with Page Blue 90 (BDH). On elution of the column with diethylamine, bands corresponding to the VHLYS mutant domains were identified, but none to the VKLYS domains. This suggested that although the mutant domains could bind to lysozyme, they could no longer associate with the VKYLS domains.
- For seven clones giving a positive reaction in ELISA, plasmids were prepared and the VKLYS gene excised by cutting with EcoRI and religating. Thus the plasmids should only direct the expression of the VHLYS mutants. 1.5 ml cultures were grown and induced for expression as above. The cells were spun down and supernatant shown to bind lysozyme as above. (Alternatively the amplified mutant VKLYS genes could have been cloned directly into the pSW1HPOLY vector for expression of the mutant activities in the absence of VKLYS.)
- An ELISA method was devised in which the activities of bacterial supernatants for binding of lysozyme (or KLH) were compared. Firstly a vector was devised for tagging of the VH domains at its C-terminal region with a peptide from the c-myc protein which is recognized by a monoclonal antibody 9E10. The vector was derived from pSW1 by a BstEII and SmaI double digest, and ligation of an oligonucleotide duplex made from
-
5′ GTC ACC GTC TCC TCA GAA CAA AAA CTC ATA TCA GAA GAG GAT CTG AAT TAA TAA 3′and 5′ TTA TTA ATT CAG ATC CTC TTC TGA GAT GAG TTT TTG TTC TGA GGA GAC G 3′. - The VHLYSMYC protein domain expressed after induction was shown to bind to lysozyme and to the 9E10 antibody by ELISA as follows:
- (1) Falcon (3912) flat bottomed wells were coated with 180 μl lysozyme (3 mg/ml) or KLH (50 μg/ml) per well in 50 mM NaHCO3, pH 9.6, and left to stand at room temperature overnight;
- (2) The wells were washed with PBS and blocked for 2 hrs at 37° C. with 200
μl 2% Sainsbury's instant dried skimmed milk powder in PBS per well; - (3) The Blocking solution was discarded, and the walls washed out with PBS (3 washes) and 150 μl test solution (supernatant or purified tagged domain) pipetted into each well. The sample was incubated at 37° C. for 2 hrs;
- (4) The test solution was discarded, and the wells washed out with PBS (3 washes). 100 μl of 4 μg/ml purified 9E10 antibody in 2% Sainsbury's instant dried skimmed milk powder in PBS was added, and incubated at 37° C. for 2 hrs;
- (5) The 9E10 antibody was discarded, the wells washed with PBS (3 washes). 100 ml of 1/500 dilution of anti-mouse antibody (peroxidase conjugated anti-mouse Ig (Dakopats)) was added and incubated at 37° C. for 2 hrs;
- (6) The second antibody was discarded and wells washed three times with PBS; and
- (7) 100
μl μl 20% hydrogen peroxide: water per 10 ml) was added to each well and the color allowed to develop for up to 10 minutes at room temperature. - The reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid, pH 4.3. ELISA plates were read in an Titertek Multiscan plate reader.
- The activities of the mutant supernatants were compared with VHLYS supernatant by competition with the VHLYSMYC domain for binding to lysozyme. The results show that supernatant from clone VHLYSMUT59 is more effective than wild type VHLYS supernatant in competing for VHLYSMYC. Furthermore, Western blots of SDS-PAGE aliquots of supernatant from the VHLYS and VHLYSMUT59 domain (using anti-Fv antisera) indicated comparable amounts of the two samples. Thus assuming identical amounts of VHLYS and VHLYSMUT59, the affinity of the mutant appears to be greater than that of the VHLYS domain.
- To check the affinity of the VHLYSMUT59 domain directly, the clone was grown at the 1 L scale and 200-300 μg purified on lysozyme-Sepharose as in Example 5. By fluorescence quench titration of samples of VHLYS and VHLYSMUT59, the number of binding sites for lysozyme were determined. The samples of VHLYS and VHLYSMUT59 were then compared in the competition ELISA with VHLYSMYC over two orders of magnitude. In the competition assay each microtitre well contained a constant amount of VHLYSMYC (approximately 0.6 μg VHLYSMYC). Varying amounts of VHLYS or VHLYSMUT59 (3.8 μM in lysozyme binding sites) were added (0.166-25 μl). The final volume and buffer concentration in all wells was constant. 9E10 (anti-myc) antibody was used to quantitate bound VHLYSMYC in each assay well. The % inhibition of VHLYSMYC binding was calculated for each addition of VHLYS or VHLYSMUT59, after subtraction of background binding. Assays were carried out in duplicate. The results indicate that VHLYSMUT59 has a higher affinity for lysozyme than VHLYS.
- The VHLYSMUT59 gene was sequenced (after recloning into M13) and shown to be identical to the VHLYS gene except for the central residues of CDR3 (Arg-Asp-Tyr-Arg). These were replaced by Thr-Gln-Arg-Pro: (encoded by ACACAAAGGCCA).
- A library of 2000 mutant VH clones was screened for lysozyme and also for KLH binding (
toothpicking 5 colonies per well as described in Example 6). Nineteen supernatants were identified with lysozyme binding activities and four with KLH binding activities. This indicates that new specificities and improved affinities can be derived by making a random repertoire of CDR3. - The finding that single domains have excellent binding activities should allow the construction of strings of domains (concatamers). Thus, multiple specificities could be built into the same molecule, allowing binding to different epitopes spaced apart by the distance between domain heads. Flexible linker regions could be built to space out the domains. In principle such molecules could be devised to have exceptional specificity and affinity.
- Two copies of the cloned heavy chain variable gene of the D1.3 antibody were linked by a nucleotide sequence encoding a flexible linker Gly-Gly-Gly-Ala-Pro-Ala-Ala-Ala-Pro-Ala-Gly-Gly-Gly- (by several steps of cutting, pasting and site directed mutagenesis) to yield the plasmid pSW3 (
FIG. 20 ). The expression was driven by a lacZ promoter and the protein was secreted into the periplasm via a pelB leader sequence (as described in Example 5 for expression of pSW1 and pSW2). The protein could be purified to homogeneity on a lysozyme affinity column. On SDS polyacrylamide gels, it gave a band of the right size (molecular weight about 26,000). The protein also bound strongly to lysozyme as detected by ELISA (see Example 5) using anti-idiotypic antiserum directed against the Fv fragment of the D1.3 antibody to detect the protein. Thus, such constructs are readily made and secreted and at least one of the domains binds to lysozyme. - A cysteine residue was introduced at the C-terminus of the VHLYS domain in the vector pSW2. The cysteine was introduced by cleavage of the vector with the restriction enzymes BstI and SmaI (which excises the C-terminal portion of the J segment) and ligation of a short oligonucleotide duplex
-
5′ GTC ACC GTC TCC TCA TGT TAA TAA 3′and 5′ TTA TTA ACA TGA GGA GAC G 3′.
By purification on an affinity column of lysozyme Sepharose it was shown that the VHLYS-Cys domain was expressed in association with the VKLYS variable domain, but the overall yields were much lower than the wild type Fv fragment. Comparison of non-reducing and reducing SDS polyacrylamide gels of the purified Fv-Cys protein indicated that the two VH-Cys domains had become linked-through the introduced cysteine residue. - Linking of enzyme activities to VH domains should be possible by either cloning the enzyme on either the N-terminal or the C-terminal side of the VH domain. Since both partners must be active, it may be necessary to design a suitable linker (see Example 8) between the two domains. For secretion of the VH-enzyme fusion, it would be preferable to utilize an enzyme which is usually secreted. In
FIGS. 21 a-21 c, there is shown the sequence of a fusion of a VH domain with alkaline phosphatase. The alkaline phosphatase gene was cloned from a plasmid carrying the E. coli alkaline phosphatase gene in a plasmid pEK48 [51] using the polymerase chain reaction. The gene was amplified with the primers -
5′CAC CAC GGT CAC CGT CTC CTC ACG GAC ACC AGA AAT GCC TGT TCT G 3′and 5′ GCG AAA ATT CAC TCC CGG GCG CGG TTT TAT TTC 3′.
The gene was introduced into the vector pSW1 by cutting at BstEII and SmaI. The construction (FIGS. 21 a-21 c) was expressed in E. coli strain BMH71-18 as in Example 5 and screened for phosphatase activity using 1 mg/ml p-nitrophenylphosphate as substrate in 10 mM diethanolamine and 0.5 mM MgCl2, pH 9.5) and also on SDS polyacrylamide gels which had been Western blotted (detecting with anti-idiotypic antiserum). No evidence was found for the secretion of the linked VHLYS-alkaline phosphatase as detected by Western blots (see Example 5), or for secretion of phosphatase activity. - However when the construct was transfected into a bacterial strain BL21DE3 [52] which is deficient in proteases, a band of the correct size (as well as degraded products) was detected on the Western blots. Furthermore phosphatase activity could now be detected in the bacterial supernatant. Such activity is not present in supernatant from the strain which had not been transfected with the construct.
- A variety of linker sequences could then be introduced at the BstEII site to improve the spacing between the two domains.
- A repertoire of Vκ genes was derived by PCR using primers as described in Example 2 from DNA prepared from mouse spleen and also from mouse spleen mRNA using the primers VK3FOR and VK2BACK and a cycle of 94° C. for 1 min, 60° C. for 1 min, 72° C. for 2 min. The PCR amplified DNA was fractionated on the agarose gel, the band excised and cloned into a vector which carries the VHLYS domain (from the D 1.3 antibody), and a cloning site (SacI and XhoI) for cloning of the light chain variable domains with a myc tail (pSW1VHLYS-VKPOLYMYC,
FIG. 22 ). - Clones were screened for lysozyme binding activities as described in Examples 5 and 7 via the myc tag on the light chain variable domain, as this should permit the following kinds of Vκ domains to be identified:
- (1) those which bind to lysozyme in the absence of the VHLYS domain;
- (2) those which associate with the heavy chain and make no contribution to binding of lysozyme; and
- (3) those which associate with the heavy chain and also contribute to binding of lysozyme (either helping or hindering).
- This would not identify those Vκ domains which associated with the VHLYS domain and completely abolished its binding to lysozyme.
- In a further experiment, the VHLYS domain was replaced by the heavy chain variable domain VH3 which had been isolated from the repertoire (see Example 6), and then the Vκ domains cloned into the vector. (Note that the VH3 domain has an internal SacI site and this was first removed to allow the cloning of the Vκ repertoire as SacI-XhoI fragments.)
- By screening the supernatant using the ELISA described in Example 6, bacterial supernatants will be identified which bind lysozyme.
- By screening several clones from a VH library derived from a mouse immunized with lysozyme via a Western blot, using the 9E10 antibody directed against the peptide tag, one clone was noted with very high levels of expression of the domain (estimated as 25-50 mg/l). The clone was sequenced to determine the nature of the sequence. The sequence proved to be closely related to that of the VHLYS domain, except with a few amino acid changes (
FIG. 23 ). The result was unexpected, and shows that a limited number of amino acid changes, perhaps even a single amino acid substitution, can cause greatly elevated levels of expression. - By making mutations of the high expressing domain at these residues, it was found that a single amino acid change in the VHLYS domain (
Asn 35 to H is) is sufficient to cause the domain to be expressed at high levels. - It can thus be seen that the present invention enables the cloning, amplification and expression of heavy and light chain variable domain encoding sequences in a much more simple manner than was previously possible. It also shows that isolated variable domains or such domains linked to effector molecules are unexpectedly useful.
- It will be appreciated that the present invention has been described above by way of example only and that variations and modifications may be made by the skilled person without departing from the scope of the invention.
-
- [1] Inbar et al., PNAS-USA, 69, 2659-2662, 1972.
- [2] Amit et al., Science, 233, 747, 1986.
- [3] Satow et al., J. Mol. Biol., 190, 593, 1986.
- [4] Colman et al., Nature, 326, 358, 1987.
- [5] Sheriff et al., PNAS-USA, 84, 8075-8079, 1987.
- [6] Padlan et al., PNAS-USA, 86, 5938-5942, 1989.
- [7] Skerra and Plückthun, Science, 240, 1038-1041, 1988.
- [8] Bird et al., Science, 242, 423-426, 1988.
- [9] Huston et al., PNAS-USA, 85, 5879-5833, 1988.
- [10] Fleischman, Arch. Biochem. Biophys. Suppl., 1, 174, 1966.
- [1,1] Porter and Weir, J. Cell. Physiol. Suppl., 1, 51, 1967.
- [1,2] Jaton et al., Biochemistry, 7, 4185, 1968.
- [1,3] Rockey, J. Exp. Med., 125, 249, 1967.
- [1,4] Stevenson, Biochem. J., 133, 827-836, 1973.
- [15] Edmundson et al., Biochemistry, 14, 3953, 1975.
- [1,6] Rossman et al., Nature, 317, 145-153, 1985.
- [1,7] Saiki et al., Science, 230, 1350-1354, 1985.
- [1,8] Larrick et al., Biochem. Biophys. Res. Comm., 160, 1250, 1989.
- [1,9] Orlandi et al., PNAS-USA, 86, 3833, 1989.
- [20] Yon and Fried, Nuc. Acids Res., 17, 4895, 1989.
- [21] Fields and Song, Nature, 340, 245-246, 1989.
- [22] Baldwin and Schultz, Science, 245, 1104-1107, 1989.
- [23] Menard et al., Cancer Res., 43, 1295-1300, 1983.
- [24] Bosslet et al., Eur. J. Nuc. Med., 14, 523-528, 1988.
- [25] Bosslet et al., Cancer Immunol. Immunother., 23, 185-191, 1986.
- [26] Bosslet et al., Int. J. Cancer, 36, 75-84, 1985.
- [27]
- [28] Bremer et al., J. Biol. Chem., 259, 14773-14777, 1984.
- [29] Griffiths & Milstein, Hybridoma Technology in the Biosciences and Medicine, 103-115, 1985.
- [30] Maniatis et al., Molecular Cloning: a Laboratory Manual, Cold Spring Harbour Laboratory, 1982.
- [31] Jones et al., Nature, 321, 522-525, 1986.
- [32] Zoller & Smith, Nuc. Acids Res., 10, 6457-6500, 1982.
- [33] Carter et al., Nuc. Acids Res., 13, 4431-4443, 1985.
- [34] Sanger et al., PNAS-USA, 74, 5463-5467, 1977.
- [35] Yannisch-Perron et al., Gene, 33, 103-119, 1985.
- [36]
- [37] Riechmann et al., Nature, 332, 323-327, 1988.
- [38] Kearney et al., J. Immunol., 123, 1548-1550, 1979.
- [39] Patter et al., PNAS-USA, 81, 7161-7163, 1984.
- [40] Galfre & Milstein, Meth. Enzym., 73, 1-46, 1981.
- [41] Laemmli, Nature, 227, 680-685, 1970.
- [42] Better et al., Science, 240, 1041, 1988.
- [43] Lei et al., J. Bacteriol., 169, 4379, 1987.
- [44] Verhoeyen et al., Science, 239, 1534, 1988.
- [45] Gronenbom, Mol. Gen. Genet, 148, 243, 1976.
- [46] Dagert et al., Gene, 6, 23, 1974.
- [47] Hanahan, J. Mol. Biol., 166, 557, 1983.
- [48] Jones et al., Nature, 321, 522, 1986.
- [49] Segal, Enzyme Kinetics, 73, Wiley, New York, 1975.
- [50] Gutfreund, Enzymes, Physical Principles, Wiley Interscience, London, 1972.
- [51] Chaidaroglou, Biochem., 27, 8338, 1988.
- [52] Grodberg and Dunn, J. Bacteriol., 170, 1245-1253, 1988.
Claims (32)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12/127,237 US20080299618A1 (en) | 1988-11-11 | 2008-05-27 | Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors |
Applications Claiming Priority (22)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB888826444A GB8826444D0 (en) | 1988-11-11 | 1988-11-11 | Cloning immunoglobulin variable domains for expression by polymerase chain reaction |
GB8826444.5 | 1988-11-11 | ||
GB8906034.7 | 1989-03-16 | ||
GB898906034A GB8906034D0 (en) | 1989-03-16 | 1989-03-16 | Recombinant dna method |
GB898909217A GB8909217D0 (en) | 1989-04-22 | 1989-04-22 | Antibody binding |
GB8909217.5 | 1989-04-22 | ||
GB8911047.2 | 1989-05-15 | ||
GB898911047A GB8911047D0 (en) | 1989-05-15 | 1989-05-15 | Antibody binding |
GB898912652A GB8912652D0 (en) | 1989-06-02 | 1989-06-02 | Antibody binding |
GB8912652.8 | 1989-06-02 | ||
GB898913900A GB8913900D0 (en) | 1989-06-16 | 1989-06-16 | Antibody binding |
GB8913900.0 | 1989-06-16 | ||
GB8918543.3 | 1989-08-15 | ||
GB898918543A GB8918543D0 (en) | 1989-08-15 | 1989-08-15 | Antibody binding |
PCT/GB1989/001344 WO1990005144A1 (en) | 1988-11-11 | 1989-11-13 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US58037490A | 1990-09-11 | 1990-09-11 | |
US79680591A | 1991-11-25 | 1991-11-25 | |
US33204694A | 1994-11-01 | 1994-11-01 | |
US08/470,031 US6248516B1 (en) | 1988-11-11 | 1995-06-06 | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US09/722,364 US6545142B1 (en) | 1988-11-11 | 2000-11-28 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,233 US20040110941A2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US12/127,237 US20080299618A1 (en) | 1988-11-11 | 2008-05-27 | Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/290,233 Continuation US20040110941A2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
Publications (1)
Publication Number | Publication Date |
---|---|
US20080299618A1 true US20080299618A1 (en) | 2008-12-04 |
Family
ID=27562806
Family Applications (5)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/470,031 Expired - Lifetime US6248516B1 (en) | 1988-11-11 | 1995-06-06 | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US09/722,364 Expired - Fee Related US6545142B1 (en) | 1988-11-11 | 2000-11-28 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,252 Expired - Fee Related US7306907B2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,233 Abandoned US20040110941A2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US12/127,237 Abandoned US20080299618A1 (en) | 1988-11-11 | 2008-05-27 | Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors |
Family Applications Before (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/470,031 Expired - Lifetime US6248516B1 (en) | 1988-11-11 | 1995-06-06 | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US09/722,364 Expired - Fee Related US6545142B1 (en) | 1988-11-11 | 2000-11-28 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,252 Expired - Fee Related US7306907B2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,233 Abandoned US20040110941A2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
Country Status (13)
Country | Link |
---|---|
US (5) | US6248516B1 (en) |
EP (1) | EP0368684B2 (en) |
JP (1) | JP2919890B2 (en) |
KR (1) | KR0184860B1 (en) |
AT (1) | ATE102631T1 (en) |
AU (1) | AU634186B2 (en) |
CA (1) | CA2002868C (en) |
DE (1) | DE68913658T3 (en) |
DK (1) | DK175392B1 (en) |
ES (1) | ES2052027T5 (en) |
FI (1) | FI903489A0 (en) |
NO (1) | NO903059L (en) |
WO (1) | WO1990005144A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8940298B2 (en) | 2007-09-04 | 2015-01-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US8940871B2 (en) | 2006-03-20 | 2015-01-27 | The Regents Of The University Of California | Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting |
Families Citing this family (1591)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5229272A (en) * | 1989-04-25 | 1993-07-20 | Igen, Inc. | Catalytic antibody components |
AU634186B2 (en) * | 1988-11-11 | 1993-02-18 | Medical Research Council | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
CA2006878A1 (en) * | 1988-12-29 | 1990-06-29 | John D. Rodwell | Molecular recognition units |
US5196510A (en) * | 1988-12-29 | 1993-03-23 | Cytogen Corporation | Molecular recognition units |
US5225538A (en) * | 1989-02-23 | 1993-07-06 | Genentech, Inc. | Lymphocyte homing receptor/immunoglobulin fusion proteins |
US6406697B1 (en) | 1989-02-23 | 2002-06-18 | Genentech, Inc. | Hybrid immunoglobulins |
US5116964A (en) * | 1989-02-23 | 1992-05-26 | Genentech, Inc. | Hybrid immunoglobulins |
DE3909799A1 (en) | 1989-03-24 | 1990-09-27 | Behringwerke Ag | MONOCLONAL ANTIBODIES (MAK) AGAINST TUMOR ASSOCIATED ANTIGENS, THEIR PRODUCTION AND USE |
US5194585A (en) * | 1989-04-25 | 1993-03-16 | Igen, Inc. | Inhibitors of catalytic antibodies |
US5599538A (en) * | 1989-04-25 | 1997-02-04 | Igen, Inc. | Autoantibodies which enhance the rate of a chemical reaction |
US6048717A (en) * | 1989-04-25 | 2000-04-11 | Igen International, Inc. | Inhibitors of catalytic antibodies |
US5318897A (en) * | 1989-04-25 | 1994-06-07 | Igen, Inc. | Monoclonal antibody and antibody components elicited to a polypeptide antigen ground state |
US5658753A (en) * | 1989-04-25 | 1997-08-19 | Paul; Sudhir | Catalytic antibody components |
US5236836A (en) * | 1989-04-25 | 1993-08-17 | Igen, Inc. | Autoantibodies which enhance the rate of a chemical reaction |
US5602015A (en) * | 1989-04-25 | 1997-02-11 | Igen, Inc. | Autoantibodies which enhance the rate of a chemical reaction |
US6291160B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6291161B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertiore |
US6291158B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
US6680192B1 (en) | 1989-05-16 | 2004-01-20 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6291159B1 (en) | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6969586B1 (en) | 1989-05-16 | 2005-11-29 | Scripps Research Institute | Method for tapping the immunological repertoire |
CA2016841C (en) * | 1989-05-16 | 1999-09-21 | William D. Huse | A method for producing polymers having a preselected activity |
US5231015A (en) * | 1989-10-18 | 1993-07-27 | Eastman Kodak Company | Methods of extracting nucleic acids and pcr amplification without using a proteolytic enzyme |
US6274324B1 (en) | 1989-12-01 | 2001-08-14 | Unilever Patent Holdings B.V. | Specific binding reagent comprising a variable domain protein linked to a support or tracer |
GB8928501D0 (en) * | 1989-12-18 | 1990-02-21 | Unilever Plc | Reagents |
ATE277179T1 (en) † | 1990-02-01 | 2004-10-15 | Dade Behring Marburg Gmbh | PRODUCTION AND USE OF HUMAN ANTIBODIES GENE BANKS (ßHUMAN ANTIBODIES LIBRARIESß) |
GB9021671D0 (en) * | 1990-10-05 | 1990-11-21 | Unilever Plc | Delivery of agents |
US5427908A (en) * | 1990-05-01 | 1995-06-27 | Affymax Technologies N.V. | Recombinant library screening methods |
US5723286A (en) * | 1990-06-20 | 1998-03-03 | Affymax Technologies N.V. | Peptide library and screening systems |
US7063943B1 (en) | 1990-07-10 | 2006-06-20 | Cambridge Antibody Technology | Methods for producing members of specific binding pairs |
GB9015198D0 (en) | 1990-07-10 | 1990-08-29 | Brien Caroline J O | Binding substance |
US6172197B1 (en) | 1991-07-10 | 2001-01-09 | Medical Research Council | Methods for producing members of specific binding pairs |
US6916605B1 (en) | 1990-07-10 | 2005-07-12 | Medical Research Council | Methods for producing members of specific binding pairs |
GB9206318D0 (en) * | 1992-03-24 | 1992-05-06 | Cambridge Antibody Tech | Binding substances |
GB9016299D0 (en) * | 1990-07-25 | 1990-09-12 | Brien Caroline J O | Binding substances |
DE4033120A1 (en) * | 1990-10-18 | 1992-04-23 | Boehringer Mannheim Gmbh | Genomic DNA fragment encoding antibody variable region prodn. - by attaching primers to hybridoma DNA then subjecting to polymerase chain reaction, for constructing genes encoding chimeric antibodies |
US5571894A (en) * | 1991-02-05 | 1996-11-05 | Ciba-Geigy Corporation | Recombinant antibodies specific for a growth factor receptor |
AU1025692A (en) * | 1991-02-06 | 1992-08-13 | Ciba-Geigy Ag | Novel chimeric antiidiotypic monoclonal antibodies |
MX9203138A (en) * | 1991-03-12 | 1992-09-01 | Biogen Inc | DOMAIN OF LINK CD2-ANTIGEN 3 (LFA-3) ASSOCIATED WITH FUNCTION LYMPHOSITES. |
JPH05508779A (en) * | 1991-03-12 | 1993-12-09 | バイオジェン,インコーポレイテッド | CD2-binding domain of lymphocyte function-related antigen 3 |
JP3672306B2 (en) * | 1991-04-10 | 2005-07-20 | ザ スクリップス リサーチ インスティテュート | Heterodimeric receptor library using phagemids |
US5858657A (en) * | 1992-05-15 | 1999-01-12 | Medical Research Council | Methods for producing members of specific binding pairs |
US6492160B1 (en) | 1991-05-15 | 2002-12-10 | Cambridge Antibody Technology Limited | Methods for producing members of specific binding pairs |
US6225447B1 (en) | 1991-05-15 | 2001-05-01 | Cambridge Antibody Technology Ltd. | Methods for producing members of specific binding pairs |
US5962255A (en) * | 1992-03-24 | 1999-10-05 | Cambridge Antibody Technology Limited | Methods for producing recombinant vectors |
US5871907A (en) * | 1991-05-15 | 1999-02-16 | Medical Research Council | Methods for producing members of specific binding pairs |
DE69233254T2 (en) | 1991-06-14 | 2004-09-16 | Genentech, Inc., South San Francisco | Humanized Heregulin antibody |
US6800738B1 (en) | 1991-06-14 | 2004-10-05 | Genentech, Inc. | Method for making humanized antibodies |
WO1994004679A1 (en) * | 1991-06-14 | 1994-03-03 | Genentech, Inc. | Method for making humanized antibodies |
US5939531A (en) * | 1991-07-15 | 1999-08-17 | Novartis Corp. | Recombinant antibodies specific for a growth factor receptor |
NL9101290A (en) * | 1991-07-23 | 1993-02-16 | Stichting Rega V Z W | RECOMBINANT DNA MOLECULA FOR THE EXPRESSION OF AN FV FRAGMENT OF AN ANTIBODY. |
US6764681B2 (en) | 1991-10-07 | 2004-07-20 | Biogen, Inc. | Method of prophylaxis or treatment of antigen presenting cell driven skin conditions using inhibitors of the CD2/LFA-3 interaction |
US5733731A (en) * | 1991-10-16 | 1998-03-31 | Affymax Technologies N.V. | Peptide library and screening method |
US5270170A (en) * | 1991-10-16 | 1993-12-14 | Affymax Technologies N.V. | Peptide library and screening method |
DK1024191T3 (en) | 1991-12-02 | 2008-12-08 | Medical Res Council | Preparation of autoantibodies displayed on phage surfaces from antibody segment libraries |
EP2224006A1 (en) * | 1991-12-02 | 2010-09-01 | MedImmune Limited | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
DE4142077A1 (en) * | 1991-12-19 | 1993-06-24 | Boehringer Mannheim Gmbh | METHOD FOR EXPRESSING RECOMBINANT ANTIKOERPERS |
US5824307A (en) | 1991-12-23 | 1998-10-20 | Medimmune, Inc. | Human-murine chimeric antibodies against respiratory syncytial virus |
US6399368B1 (en) | 1992-01-17 | 2002-06-04 | Board Of Regents, The University Of Texas System | Secretion of T cell receptor fragments from recombinant Escherichia coli cells |
CZ291039B6 (en) | 1992-02-06 | 2002-12-11 | Schering Corporation | Monoclonal antibody, hybridoma, polypeptide and process for preparing thereof, isolated DNA, recombinant vector, host cell, humanized antibody and pharmaceutical composition |
US5733743A (en) * | 1992-03-24 | 1998-03-31 | Cambridge Antibody Technology Limited | Methods for producing members of specific binding pairs |
EP0640094A1 (en) * | 1992-04-24 | 1995-03-01 | The Board Of Regents, The University Of Texas System | Recombinant production of immunoglobulin-like domains in prokaryotic cells |
GB9216983D0 (en) * | 1992-08-11 | 1992-09-23 | Unilever Plc | Polypeptide production |
US6765087B1 (en) | 1992-08-21 | 2004-07-20 | Vrije Universiteit Brussel | Immunoglobulins devoid of light chains |
DE4233152A1 (en) * | 1992-10-02 | 1994-04-07 | Behringwerke Ag | Antibody-enzyme conjugates for prodrug activation |
GB9225453D0 (en) | 1992-12-04 | 1993-01-27 | Medical Res Council | Binding proteins |
DK1231268T3 (en) * | 1994-01-31 | 2005-11-21 | Univ Boston | Polyclonal antibody libraries |
US6010861A (en) * | 1994-08-03 | 2000-01-04 | Dgi Biotechnologies, Llc | Target specific screens and their use for discovering small organic molecular pharmacophores |
US6056957A (en) * | 1994-08-04 | 2000-05-02 | Schering Corporation | Humanized monoclonal antibodies against human interleukin-5 |
EP0859841B1 (en) | 1995-08-18 | 2002-06-19 | MorphoSys AG | Protein/(poly)peptide libraries |
US6828422B1 (en) | 1995-08-18 | 2004-12-07 | Morphosys Ag | Protein/(poly)peptide libraries |
US7368111B2 (en) | 1995-10-06 | 2008-05-06 | Cambridge Antibody Technology Limited | Human antibodies specific for TGFβ2 |
US6136311A (en) | 1996-05-06 | 2000-10-24 | Cornell Research Foundation, Inc. | Treatment and diagnosis of cancer |
DE69738254T2 (en) | 1996-05-10 | 2008-08-14 | Novozymes A/S | METHOD FOR PROVISION OF DNA SEQUENCES |
ATE391183T1 (en) | 1996-08-19 | 2008-04-15 | Morphosys Ip Gmbh | VECTORS/DNA SEQUENCES FROM HUMAN COMBINATORY ANTIBODIES LIBRARIES |
GB9701425D0 (en) | 1997-01-24 | 1997-03-12 | Bioinvent Int Ab | A method for in vitro molecular evolution of protein function |
ATE461282T1 (en) * | 1997-10-27 | 2010-04-15 | Bac Ip Bv | MULTIVALENT ANTIGEN-BINDING PROTEINS |
WO1999040434A1 (en) | 1998-02-04 | 1999-08-12 | Invitrogen Corporation | Microarrays and uses therefor |
CA2328422A1 (en) | 1998-05-13 | 1999-11-18 | Diversys Limited | Selection system |
US6914128B1 (en) | 1999-03-25 | 2005-07-05 | Abbott Gmbh & Co. Kg | Human antibodies that bind human IL-12 and methods for producing |
US6492497B1 (en) | 1999-04-30 | 2002-12-10 | Cambridge Antibody Technology Limited | Specific binding members for TGFbeta1 |
US7534605B2 (en) | 1999-06-08 | 2009-05-19 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | CD44 polypeptides, polynucleotides encoding same, antibodies directed thereagainst and method of using same for diagnosing and treating inflammatory diseases |
US7297478B1 (en) | 2000-09-22 | 2007-11-20 | Large Scale Biology Corporation | Creation of variable length and sequence linker regions for dual-domain or multi-domain molecules |
AU785038B2 (en) | 2000-01-27 | 2006-08-31 | Applied Molecular Evolution, Inc. | Ultra high affinity neutralizing antibodies |
AU2001234125A1 (en) | 2000-02-22 | 2001-09-03 | Medical And Biological Laboratories Co., Ltd. | Antibody library |
EP2341074A1 (en) | 2000-03-01 | 2011-07-06 | MedImmune, LLC | Antibodies binding to the f protein of a respiratory syncytial virus (rsv) |
US8288322B2 (en) | 2000-04-17 | 2012-10-16 | Dyax Corp. | Methods of constructing libraries comprising displayed and/or expressed members of a diverse family of peptides, polypeptides or proteins and the novel libraries |
CA2406236C (en) * | 2000-04-17 | 2013-02-19 | Dyax Corp. | Novel methods of constructing libraries of genetic packages that collectively display the members of a diverse family of peptides, polypeptides or proteins |
EP1176200A3 (en) | 2000-06-20 | 2005-01-12 | Switch Biotech Aktiengesellschaft | Use of polyeptides or their encoding nucleic acids for the diagnosis or treatment of skin diseases or wound healing and their use in indentifying pharmacologically acitve substances |
ATE420958T1 (en) | 2000-06-29 | 2009-01-15 | Abbott Lab | ANTIBODIES WITH DUAL SPECIFICITIES AND METHOD FOR THE PRODUCTION AND USE THEREOF |
US7288390B2 (en) | 2000-08-07 | 2007-10-30 | Centocor, Inc. | Anti-dual integrin antibodies, compositions, methods and uses |
US6902734B2 (en) | 2000-08-07 | 2005-06-07 | Centocor, Inc. | Anti-IL-12 antibodies and compositions thereof |
UA81743C2 (en) | 2000-08-07 | 2008-02-11 | Центокор, Инк. | HUMAN MONOCLONAL ANTIBODY WHICH SPECIFICALLY BINDS TUMOR NECROSIS FACTOR ALFA (TNFα), PHARMACEUTICAL MIXTURE CONTAINING THEREOF, AND METHOD FOR TREATING ARTHRITIS |
US20050196755A1 (en) * | 2000-11-17 | 2005-09-08 | Maurice Zauderer | In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells |
AU2001297872B2 (en) | 2000-11-17 | 2006-11-09 | University Of Rochester | In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells |
US7179900B2 (en) | 2000-11-28 | 2007-02-20 | Medimmune, Inc. | Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment |
US6919189B2 (en) | 2000-12-11 | 2005-07-19 | Alexion Pharmaceuticals, Inc. | Nested oligonucleotides containing a hairpin for nucleic acid amplification |
US6958213B2 (en) | 2000-12-12 | 2005-10-25 | Alligator Bioscience Ab | Method for in vitro molecular evolution of protein function |
US7658921B2 (en) | 2000-12-12 | 2010-02-09 | Medimmune, Llc | Molecules with extended half-lives, compositions and uses thereof |
EP2341060B1 (en) | 2000-12-12 | 2019-02-20 | MedImmune, LLC | Molecules with extended half-lives, compositions and uses thereof |
AU2002249854B2 (en) | 2000-12-18 | 2007-09-20 | Dyax Corp. | Focused libraries of genetic packages |
US20020086292A1 (en) | 2000-12-22 | 2002-07-04 | Shigeaki Harayama | Synthesis of hybrid polynucleotide molecules using single-stranded polynucleotide molecules |
WO2002051438A2 (en) | 2000-12-22 | 2002-07-04 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Use of repulsive guidance molecule (rgm) and its modulators |
WO2002086096A2 (en) * | 2001-01-23 | 2002-10-31 | University Of Rochester Medical Center | Methods of producing or identifying intrabodies in eukaryotic cells |
JP2005503999A (en) | 2001-01-31 | 2005-02-10 | アイデック ファーマスーティカルズ コーポレイション | Use of CD23 antagonists for the treatment of neoplastic diseases |
GB0110029D0 (en) | 2001-04-24 | 2001-06-13 | Grosveld Frank | Transgenic animal |
US6972324B2 (en) | 2001-05-18 | 2005-12-06 | Boehringer Ingelheim Pharmaceuticals, Inc. | Antibodies specific for CD44v6 |
GB0115841D0 (en) * | 2001-06-28 | 2001-08-22 | Medical Res Council | Ligand |
US20060073141A1 (en) * | 2001-06-28 | 2006-04-06 | Domantis Limited | Compositions and methods for treating inflammatory disorders |
ATE477280T1 (en) * | 2001-06-28 | 2010-08-15 | Domantis Ltd | DOUBLE-SPECIFIC LIGAND AND USE THEREOF |
WO2004003019A2 (en) * | 2002-06-28 | 2004-01-08 | Domantis Limited | Immunoglobin single variant antigen-binding domains and dual-specific constructs |
WO2003009740A2 (en) | 2001-07-24 | 2003-02-06 | Biogen Idec Ma Inc. | Methods for treating or preventing sclerotic disorders using cd2-binding agents |
US6833441B2 (en) | 2001-08-01 | 2004-12-21 | Abmaxis, Inc. | Compositions and methods for generating chimeric heteromultimers |
EP2202243A3 (en) | 2001-08-10 | 2012-08-08 | Aberdeen University | Antigen binding domains from fish |
WO2003018749A2 (en) * | 2001-08-22 | 2003-03-06 | Shengfeng Li | Compositions and methods for generating antigen-binding units |
US20040005709A1 (en) * | 2001-10-24 | 2004-01-08 | Hoogenboom Henricus Renerus Jacobus Mattheus | Hybridization control of sequence variation |
GB0126378D0 (en) | 2001-11-02 | 2002-01-02 | Oxford Biomedica Ltd | Antigen |
US7175983B2 (en) | 2001-11-02 | 2007-02-13 | Abmaxis, Inc. | Adapter-directed display systems |
CA2471116A1 (en) * | 2001-12-21 | 2003-07-03 | Serge Muyldermans | Method for cloning of variable domain sequences |
US20050069549A1 (en) | 2002-01-14 | 2005-03-31 | William Herman | Targeted ligands |
SI2508596T1 (en) | 2002-02-21 | 2016-01-29 | Institute Of Virology Slovak Academy Of Sciences | MN/CA IX-specific monoclonal antibodies generated from MN/CA IX-deficient mice and methods of use |
US7718776B2 (en) * | 2002-04-05 | 2010-05-18 | Amgen Inc. | Human anti-OPGL neutralizing antibodies as selective OPGL pathway inhibitors |
US7135310B2 (en) | 2002-04-24 | 2006-11-14 | The Regents Of The University Of California | Method to amplify variable sequences without imposing primer sequences |
ES2656427T3 (en) | 2002-05-22 | 2018-02-27 | Esbatech, An Alcon Biomedical Research Unit Llc | Immunoglobulin frames demonstrating improved stability in the intracellular environment and methods for identification |
EP2305710A3 (en) | 2002-06-03 | 2013-05-29 | Genentech, Inc. | Synthetic antibody phage libraries |
WO2003102157A2 (en) | 2002-06-03 | 2003-12-11 | Genentech, Inc. | Synthetic antibody phage libraries |
US7425618B2 (en) | 2002-06-14 | 2008-09-16 | Medimmune, Inc. | Stabilized anti-respiratory syncytial virus (RSV) antibody formulations |
GB0213745D0 (en) | 2002-06-14 | 2002-07-24 | Univ Edinburgh | Enzyme |
US7132100B2 (en) | 2002-06-14 | 2006-11-07 | Medimmune, Inc. | Stabilized liquid anti-RSV antibody formulations |
US9028822B2 (en) | 2002-06-28 | 2015-05-12 | Domantis Limited | Antagonists against TNFR1 and methods of use therefor |
US20060002935A1 (en) | 2002-06-28 | 2006-01-05 | Domantis Limited | Tumor Necrosis Factor Receptor 1 antagonists and methods of use therefor |
US9321832B2 (en) | 2002-06-28 | 2016-04-26 | Domantis Limited | Ligand |
US7696320B2 (en) | 2004-08-24 | 2010-04-13 | Domantis Limited | Ligands that have binding specificity for VEGF and/or EGFR and methods of use therefor |
WO2004015425A1 (en) | 2002-08-07 | 2004-02-19 | Umc Utrecht Holding B.V. | Modulation of platelet adhesion based on the surface exposed beta-switch loop of platelet glycoprotein ib-alpha |
US20040067532A1 (en) | 2002-08-12 | 2004-04-08 | Genetastix Corporation | High throughput generation and affinity maturation of humanized antibody |
NZ593428A (en) * | 2002-09-06 | 2013-01-25 | Amgen Inc | Therapeutic human anti-il-1r1 monoclonal antibody |
EP1947113B1 (en) | 2002-10-07 | 2011-12-14 | Ludwig Institute for Cancer Research Ltd | P53 binding polypeptide |
WO2004034988A2 (en) * | 2002-10-16 | 2004-04-29 | Amgen Inc. | Human anti-ifn-ϝ neutralizing antibodies as selective ifn-ϝ pathway inhibitors |
US9701754B1 (en) | 2002-10-23 | 2017-07-11 | City Of Hope | Covalent disulfide-linked diabodies and uses thereof |
US20060034845A1 (en) | 2002-11-08 | 2006-02-16 | Karen Silence | Single domain antibodies directed against tumor necrosis factor alpha and uses therefor |
NZ540194A (en) | 2002-11-08 | 2008-07-31 | Ablynx Nv | Single domain antibodies directed against tumour necrosis factor-alpha and uses therefor |
US9320792B2 (en) | 2002-11-08 | 2016-04-26 | Ablynx N.V. | Pulmonary administration of immunoglobulin single variable domains and constructs thereof |
EP1578801A2 (en) * | 2002-12-27 | 2005-09-28 | Domantis Limited | Dual specific single domain antibodies specific for a ligand and for the receptor of the ligand |
GB0230201D0 (en) * | 2002-12-27 | 2003-02-05 | Domantis Ltd | Retargeting |
GB0230203D0 (en) * | 2002-12-27 | 2003-02-05 | Domantis Ltd | Fc fusion |
ES2542330T3 (en) | 2003-01-10 | 2015-08-04 | Ablynx N.V. | Therapeutic polypeptides, homologs thereof, fragments thereof and their use in modulating platelet-mediated aggregation |
EP1590369B1 (en) * | 2003-01-24 | 2016-03-16 | Applied Molecular Evolution, Inc. | Human il-1 beta antagonists |
DE10303974A1 (en) | 2003-01-31 | 2004-08-05 | Abbott Gmbh & Co. Kg | Amyloid β (1-42) oligomers, process for their preparation and their use |
SI1606409T1 (en) | 2003-03-19 | 2011-01-31 | Biogen Idec Inc | Nogo receptor binding protein |
TWI353991B (en) | 2003-05-06 | 2011-12-11 | Syntonix Pharmaceuticals Inc | Immunoglobulin chimeric monomer-dimer hybrids |
US9708410B2 (en) | 2003-05-30 | 2017-07-18 | Janssen Biotech, Inc. | Anti-tissue factor antibodies and compositions |
EP1498133A1 (en) | 2003-07-18 | 2005-01-19 | Aventis Pharma Deutschland GmbH | Use of a pak inhibitor for the treatment of a joint disease |
US20050106667A1 (en) | 2003-08-01 | 2005-05-19 | Genentech, Inc | Binding polypeptides with restricted diversity sequences |
JP2007501011A (en) * | 2003-08-01 | 2007-01-25 | ジェネンテック・インコーポレーテッド | Binding polypeptide having restriction diversity sequence |
US7758859B2 (en) | 2003-08-01 | 2010-07-20 | Genentech, Inc. | Anti-VEGF antibodies |
EP2824190A1 (en) * | 2003-09-09 | 2015-01-14 | Integrigen, Inc. | Methods and compositions for generation of germline human antibody genes |
ES2339710T5 (en) | 2003-09-23 | 2017-10-05 | University Of North Carolina At Chapel Hill | Cells that coexpress vitamin K reductase and vitamin K dependent protein and use them to improve the productivity of said vitamin K dependent protein |
WO2006101474A1 (en) | 2005-03-15 | 2006-09-28 | University Of North Carolina At Chapel Hill | Methods and compositions for producing active vitamin k-dependent proteins |
ES2344413T3 (en) | 2003-10-14 | 2010-08-26 | Baxter International Inc. | VKORC1 POLYPEPTIDE FOR RECYCLING VITAMIN K-EPOXIDE, A THERAPEUTIC TARGET OF CUMARINE AND ITS DERIVATIVES. |
WO2005047327A2 (en) | 2003-11-12 | 2005-05-26 | Biogen Idec Ma Inc. | NEONATAL Fc RECEPTOR (FcRn)-BINDING POLYPEPTIDE VARIANTS, DIMERIC Fc BINDING PROTEINS AND METHODS RELATED THERETO |
WO2005054860A1 (en) | 2003-12-01 | 2005-06-16 | Dako Denmark A/S | Methods and compositions for immuno-histochemical detection |
GB0328690D0 (en) | 2003-12-10 | 2004-01-14 | Ludwig Inst Cancer Res | Tumour suppressor protein |
PL2805728T3 (en) | 2003-12-23 | 2020-07-13 | Genentech, Inc. | Novel anti-IL 13 antibodies and uses thereof |
EP1761561B1 (en) * | 2004-01-20 | 2015-08-26 | KaloBios Pharmaceuticals, Inc. | Antibody specificity transfer using minimal essential binding determinants |
GB0406215D0 (en) | 2004-03-19 | 2004-04-21 | Procure Therapeutics Ltd | Prostate stem cell |
NZ550225A (en) | 2004-03-30 | 2010-11-26 | Glaxo Group Ltd | Immunoglobulins that bind oncostatin and inhibit or block interaction between hOSM and pg130 |
US7785903B2 (en) | 2004-04-09 | 2010-08-31 | Genentech, Inc. | Variable domain library and uses |
ES2442386T3 (en) | 2004-04-23 | 2014-02-11 | Bundesrepublik Deutschland Letztvertreten Durch Das Robert Koch-Institut Vertreten Durch Seinen Pr | Method for the treatment of conditions mediated by T cells by the decrease of positive ICOS cells in vivo. |
US7662921B2 (en) | 2004-05-07 | 2010-02-16 | Astellas Us Llc | Methods of treating viral disorders |
MXPA06014031A (en) | 2004-06-01 | 2007-10-08 | Domantis Ltd | Drug compositions, fusions and conjugates. |
CA2567655C (en) | 2004-06-02 | 2015-06-30 | Diatech Pty Ltd | Binding moieties based on shark ignar domains |
EP1776136B1 (en) | 2004-06-24 | 2012-10-03 | Biogen Idec MA Inc. | Treatment of conditions involving demyelination |
TWI307630B (en) | 2004-07-01 | 2009-03-21 | Glaxo Group Ltd | Immunoglobulins |
GB0414886D0 (en) | 2004-07-02 | 2004-08-04 | Neutec Pharma Plc | Treatment of bacterial infections |
PL2053408T3 (en) | 2004-07-20 | 2012-08-31 | Symphogen As | A procedure for structural characterization of a recombinant polyclonal protein or a polyclonal cell line |
KR20070038556A (en) | 2004-07-20 | 2007-04-10 | 심포젠 에이/에스 | Anti-rhesus d recombinant polyclonal antibody and methods of manufacture |
PL2311874T3 (en) | 2004-07-22 | 2017-10-31 | Univ Erasmus Med Ct Rotterdam | Binding molecules |
GB0416487D0 (en) | 2004-07-23 | 2004-08-25 | Isis Innovation | Modified virus |
CN101014245A (en) | 2004-08-03 | 2007-08-08 | 比奥根艾迪克Ma公司 | Taj in neuronal function |
US7563443B2 (en) | 2004-09-17 | 2009-07-21 | Domantis Limited | Monovalent anti-CD40L antibody polypeptides and compositions thereof |
ES2429541T3 (en) | 2004-11-16 | 2013-11-15 | Kalobios Pharmaceuticals, Inc. | Immunoglobulin variable region cassette exchange |
GB0425739D0 (en) * | 2004-11-23 | 2004-12-22 | Procure Therapeutics Ltd | Humanised baculovirus 2 |
GB0521621D0 (en) | 2005-10-24 | 2005-11-30 | Domantis Ltd | Tumor necrosis factor receptor 1 antagonists for treating respiratory diseases |
FR2879605B1 (en) | 2004-12-16 | 2008-10-17 | Centre Nat Rech Scient Cnrse | PRODUCTION OF ANTIBODY FORMATS AND IMMUNOLOGICAL APPLICATIONS OF THESE FORMATS |
WO2006071200A2 (en) | 2004-12-30 | 2006-07-06 | Agency For Science, Technology And Research | Chinese hamster apoptosis-related genes |
AU2006203889A1 (en) | 2005-01-05 | 2006-07-13 | Biogen Idec Ma Inc. | CRIPTO binding molecules |
PT1836500E (en) | 2005-01-14 | 2010-09-28 | Ablynx Nv | Methods and assays for distinguishing between different forms of diseases and disorders characterized by thrombocytopenia and/or by spontaneous interaction between von willebrand factor (vwf) and platelets |
AU2006222204B2 (en) | 2005-03-11 | 2012-09-27 | Sanofi-Aventis | Use of MGC4504 |
JP2008532559A (en) | 2005-03-19 | 2008-08-21 | メディカル リサーチ カウンシル | Treatment and prevention of viral infection or improvement of treatment and prevention |
PT1866339E (en) | 2005-03-25 | 2013-09-03 | Gitr Inc | Gitr binding molecules and uses therefor |
JP2008539742A (en) | 2005-05-11 | 2008-11-20 | サノフィ−アベンティス | Use of GIP promoter polymorphism |
HUE045710T2 (en) | 2005-05-18 | 2020-01-28 | Ablynx Nv | Improved nanobodies tm against tumor necrosis factor-alpha |
PE20061444A1 (en) | 2005-05-19 | 2007-01-15 | Centocor Inc | ANTI-MCP-1 ANTIBODY, COMPOSITIONS, METHODS AND USES |
EP3415535B1 (en) | 2005-05-20 | 2020-12-09 | Ablynx N.V. | Improved nanobodies tm for the treatment of aggregation-mediated disorders |
JP5372500B2 (en) | 2005-06-17 | 2013-12-18 | トレラクス リクイデーティング トラスト | ILT3-binding molecules and uses thereof |
KR20130080058A (en) | 2005-06-30 | 2013-07-11 | 아보트 러보러터리즈 | Il-12/p40 binding proteins |
EP2478917A1 (en) | 2005-07-08 | 2012-07-25 | Biogen Idec MA Inc. | SP35 antibodies and uses thereof |
EP2500352A1 (en) | 2005-08-19 | 2012-09-19 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
US7612181B2 (en) | 2005-08-19 | 2009-11-03 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
EP2500356A3 (en) | 2005-08-19 | 2012-10-24 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
RS55788B1 (en) | 2005-08-31 | 2017-08-31 | Merck Sharp & Dohme | Engineered anti-il-23 antibodies |
CN101277974A (en) | 2005-09-30 | 2008-10-01 | 阿伯特有限及两合公司 | Binding domains of proteins of the repulsive guidance molecule (RGM) protein family and functional fragments thereof, and their use |
WO2007041644A1 (en) | 2005-10-03 | 2007-04-12 | Smith & Nephew, Inc. | Locking instrument assembly |
DE102005048898A1 (en) | 2005-10-12 | 2007-04-19 | Sanofi-Aventis Deutschland Gmbh | EGLN2 variants and their use in the prevention or treatment of thromboembolic disorders and coronary heart disease |
GB0521139D0 (en) | 2005-10-18 | 2005-11-23 | Univ Sheffield | Therapeutic agent |
WO2007053524A2 (en) * | 2005-10-28 | 2007-05-10 | The Florida International University Board Of Trustees | Horse: human chimeric antibodies |
US8753625B2 (en) | 2005-11-04 | 2014-06-17 | Genentech, Inc. | Use of complement inhibitors to treat ocular diseases |
US20090246189A1 (en) | 2005-11-04 | 2009-10-01 | Biogen Idec Ma Inc. And Mclean Hospital | Methods for Promoting Neurite Outgrowth and Survival of Dopaminergic Neurons |
ES2577292T3 (en) | 2005-11-07 | 2016-07-14 | Genentech, Inc. | Binding polypeptides with diversified VH / VL hypervariable sequences and consensus |
UA96139C2 (en) | 2005-11-08 | 2011-10-10 | Дженентек, Інк. | Anti-neuropilin-1 (nrp1) antibody |
JP2009516513A (en) | 2005-11-21 | 2009-04-23 | ラボラトワール セローノ ソシエテ アノニム | Composition and production method of hybrid antigen binding molecule and use thereof |
SG10201706600VA (en) | 2005-11-30 | 2017-09-28 | Abbvie Inc | Monoclonal antibodies and uses thereof |
PT1954718E (en) | 2005-11-30 | 2014-12-16 | Abbvie Inc | Anti-a globulomer antibodies, antigen-binding moieties thereof, corresponding hybridomas, nucleic acids, vectors, host cells, methods of producing said antibodies, compositions comprising said antibodies, uses of said antibodies and methods of using said antibodies |
AU2006321364B2 (en) * | 2005-12-01 | 2011-11-10 | Domantis Limited | Noncompetitive domain antibody formats that bind Interleukin 1 Receptor type 1 |
ES2547689T3 (en) | 2005-12-02 | 2015-10-08 | Genentech, Inc. | Compositions and methods for the treatment of diseases and disorders associated with cytokine signaling that involve antibodies that bind to IL-22 and IL-22R |
US20090175872A1 (en) | 2005-12-02 | 2009-07-09 | Biogen Idec Ma Inc. | Treatment of Conditions Involving Demyelination |
EP1973951A2 (en) * | 2005-12-02 | 2008-10-01 | Genentech, Inc. | Binding polypeptides with restricted diversity sequences |
RU2470941C2 (en) | 2005-12-02 | 2012-12-27 | Дженентек, Инк. | Binding polypeptides and use thereof |
EP1981902B1 (en) | 2006-01-27 | 2015-07-29 | Biogen MA Inc. | Nogo receptor antagonists |
EA017417B1 (en) | 2006-02-01 | 2012-12-28 | Сефалон Астралия Пти Лтд. | DOMAIN ANTIBODY CONSTRUCT WHICH BINDS TO HUMAN TNF-α AND USE THEREOF |
WO2007100711A2 (en) | 2006-02-24 | 2007-09-07 | Investigen, Inc. | Methods and compositions for detecting polynucleotides |
US20100226920A1 (en) * | 2006-03-27 | 2010-09-09 | Ablynx N.V. | Medical delivery device for therapeutic proteins based on single domain antibodies |
EP2016101A2 (en) * | 2006-05-09 | 2009-01-21 | Genentech, Inc. | Binding polypeptides with optimized scaffolds |
KR101454508B1 (en) | 2006-05-30 | 2014-11-04 | 제넨테크, 인크. | Antibodies and immunoconjugates and uses therefor |
GB0611116D0 (en) | 2006-06-06 | 2006-07-19 | Oxford Genome Sciences Uk Ltd | Proteins |
CA2655903A1 (en) | 2006-06-19 | 2008-08-07 | Tolerx, Inc. | Ilt3 binding molecules and uses therefor |
US8874380B2 (en) | 2010-12-09 | 2014-10-28 | Rutgers, The State University Of New Jersey | Method of overcoming therapeutic limitations of nonuniform distribution of radiopharmaceuticals and chemotherapy drugs |
WO2008019061A2 (en) * | 2006-08-03 | 2008-02-14 | Vaccinex, Inc. | Anti-il-6 monoclonal antibodies and uses thereof |
DK2059533T3 (en) | 2006-08-30 | 2013-02-25 | Genentech Inc | MULTI-SPECIFIC ANTIBODIES |
WO2008030611A2 (en) | 2006-09-05 | 2008-03-13 | Medarex, Inc. | Antibodies to bone morphogenic proteins and receptors therefor and methods for their use |
WO2008127271A2 (en) | 2006-09-08 | 2008-10-23 | Abbott Laboratories | Interleukin -13 binding proteins |
JP2010503407A (en) | 2006-09-12 | 2010-02-04 | ジェネンテック・インコーポレーテッド | Methods and compositions for diagnosis and treatment of cancer |
US7833527B2 (en) | 2006-10-02 | 2010-11-16 | Amgen Inc. | Methods of treating psoriasis using IL-17 Receptor A antibodies |
KR20170123712A (en) | 2006-10-02 | 2017-11-08 | 메다렉스, 엘.엘.시. | Human antibodies that bind cxcr4 and uses thereof |
GB0620705D0 (en) | 2006-10-18 | 2006-11-29 | Opsona Therapeutics | Compounds for the modulation of toll-like receptor activity and assay methods for the identification of said compounds |
WO2008052187A2 (en) | 2006-10-27 | 2008-05-02 | Genentech. Inc. | Antibodies and immunoconjugates and uses therefor |
US8067179B2 (en) | 2006-11-30 | 2011-11-29 | Research Development Foundation | Immunoglobulin libraries |
US8455626B2 (en) | 2006-11-30 | 2013-06-04 | Abbott Laboratories | Aβ conformer selective anti-aβ globulomer monoclonal antibodies |
BRPI0717902A2 (en) | 2006-12-01 | 2013-10-29 | Medarex Inc | "HUMAN MONOCLONAL ANTIBODY ISOLATED, COMPOSITION, ASSOCIATED ANTIBODY-MOLLECLE PARTNERSHIP, IMMUNOCOUGHTED, ISOLATED NUCLEIC ACID MOLECULES, EXPRESSION VECTOR, HOSPEDIC CELL FOR PREPARING A CDT FOR THE PREPARATION OF A CD22 AND METHOD FOR TREATING INFLAMMATORY DISEASE OR SELF-IMMUNEING AN INDIVIDUAL " |
CL2007003622A1 (en) | 2006-12-13 | 2009-08-07 | Medarex Inc | Human anti-cd19 monoclonal antibody; composition comprising it; and tumor cell growth inhibition method. |
BRPI0720271A2 (en) | 2006-12-14 | 2014-01-28 | Schering Corp | DESIGNED ANTI-TSLP ANTIBODY |
NZ578354A (en) | 2006-12-14 | 2012-01-12 | Medarex Inc | Antibody-partner molecule conjugates that bind cd70 and uses thereof |
CA2670992C (en) | 2006-12-18 | 2017-11-21 | Genentech, Inc. | Antagonist anti-notch3 antibodies and their use in the prevention and treatment of notch3-related diseases |
US9512236B2 (en) | 2006-12-19 | 2016-12-06 | Ablynx N.V. | Amino acid sequences directed against GPCRS and polypeptides comprising the same for the treatment of GPCR-related diseases and disorders |
CA2673331A1 (en) | 2006-12-19 | 2008-06-26 | Ablynx N.V. | Amino acid sequences directed against gpcrs and polypeptides comprising the same for the treatment of gpcr-related diseases and disorders |
AU2007336243B2 (en) | 2006-12-19 | 2012-07-26 | Ablynx N.V. | Amino acid sequences directed against a metalloproteinase from the ADAM family and polypeptides comprising the same for the treatment of ADAM-related diseases and disorders |
US8128926B2 (en) | 2007-01-09 | 2012-03-06 | Biogen Idec Ma Inc. | Sp35 antibodies and uses thereof |
GEP20125693B (en) | 2007-01-09 | 2012-11-26 | Biogen Idec Inc | Sp35 antibodies and usage thereof |
AU2008205512B2 (en) | 2007-01-16 | 2014-06-12 | Abbvie Inc. | Methods for treating psoriasis |
US8664364B2 (en) | 2007-01-24 | 2014-03-04 | Carnegie Mellon University | Optical biosensors |
US7771947B2 (en) | 2007-02-23 | 2010-08-10 | Investigen, Inc. | Methods and compositions for rapid light-activated isolation and detection of analytes |
EP2064242A1 (en) | 2007-02-23 | 2009-06-03 | Schering Corporation | Engineered anti-il-23p19 antibodies |
PL2059534T3 (en) | 2007-02-23 | 2012-09-28 | Merck Sharp & Dohme | Engineered anti-il-23p19 antibodies |
EP3118221B1 (en) | 2007-02-26 | 2019-08-21 | Oxford BioTherapeutics Ltd | Proteins |
WO2008104803A2 (en) | 2007-02-26 | 2008-09-04 | Oxford Genome Sciences (Uk) Limited | Proteins |
US20100311767A1 (en) | 2007-02-27 | 2010-12-09 | Abbott Gmbh & Co. Kg | Method for the treatment of amyloidoses |
AU2008219684B2 (en) | 2007-02-28 | 2014-04-17 | Merck Sharp & Dohme Corp. | Engineered anti-IL-23R antibodies |
RU2518340C2 (en) | 2007-03-30 | 2014-06-10 | Эббви Инк | Recombinant expression vector elements (reves) to enhance expression of recombinant proteins in host cells |
US9969797B2 (en) * | 2008-04-23 | 2018-05-15 | Covalent Bioscience Incorporated | Immunoglobulins directed to bacterial, viral and endogenous polypeptides |
EP2139916A1 (en) | 2007-04-26 | 2010-01-06 | Opsona Therapeutics Limited | Toll-like receptor binding epitope and compositions for binding thereto |
TWI570135B (en) | 2007-04-27 | 2017-02-11 | 建南德克公司 | Potent, stable and non-immunosuppressive anti-cd4 antibodies |
DK2155789T3 (en) | 2007-05-01 | 2013-10-21 | Res Dev Foundation | Immunoglobulin Fc libraries |
WO2008143666A2 (en) | 2007-05-17 | 2008-11-27 | Genentech, Inc. | Crystal structures of neuropilin fragments and neuropilin-antibody complexes |
EP1997830A1 (en) | 2007-06-01 | 2008-12-03 | AIMM Therapeutics B.V. | RSV specific binding molecules and means for producing them |
CA2683801A1 (en) * | 2007-06-06 | 2008-12-11 | Domantis Limited | Polypeptides, antibody variable domains and antagonists |
MX2009013137A (en) | 2007-06-06 | 2010-04-30 | Domantis Ltd | Methods for selecting protease resistant polypeptides. |
GB0724331D0 (en) | 2007-12-13 | 2008-01-23 | Domantis Ltd | Compositions for pulmonary delivery |
US8138313B2 (en) | 2007-06-15 | 2012-03-20 | Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts | Treatment of tumors using specific anti-L1 antibody |
WO2009002451A2 (en) | 2007-06-22 | 2008-12-31 | Genera, Doo | Adamts4 as a blood biomarker and therapeutic target for chronis renal failure |
WO2009004066A2 (en) | 2007-07-03 | 2009-01-08 | Ablynx N.V. | Providing improved immunoglobulin sequences by mutating cdr and/or fr positions |
CN101801413A (en) | 2007-07-12 | 2010-08-11 | 托勒克斯股份有限公司 | Combination therapies employing GITR binding molecules |
EP2023144A1 (en) | 2007-08-01 | 2009-02-11 | Sanofi-Aventis | Novel AS160-like protein, test systems, methods and uses involving it for the identification of diabetes type 2 therapeutics |
AU2008285626B2 (en) | 2007-08-03 | 2013-09-12 | Neuramedy Co., Ltd. | Use of TLR-2 antagonists for treatment of reperfusion injury and tissue damage |
NO2195023T3 (en) * | 2007-08-29 | 2018-08-04 | ||
MX2010002661A (en) | 2007-09-14 | 2010-05-20 | Adimab Inc | Rationally designed, synthetic antibody libraries and uses therefor. |
US8877688B2 (en) | 2007-09-14 | 2014-11-04 | Adimab, Llc | Rationally designed, synthetic antibody libraries and uses therefor |
CA2698390C (en) | 2007-09-18 | 2018-05-22 | Dako Denmark A/S | A rapid and sensitive method for detection of biological targets |
DK3059246T3 (en) | 2007-09-26 | 2018-10-01 | Chugai Pharmaceutical Co Ltd | Modified constant region of an antibody |
CN104004088B (en) | 2007-09-26 | 2017-11-07 | Ucb医药有限公司 | dual specificity antibody fusions |
EP2050764A1 (en) * | 2007-10-15 | 2009-04-22 | sanofi-aventis | Novel polyvalent bispecific antibody format and uses thereof |
EP3360567A1 (en) | 2007-11-07 | 2018-08-15 | Genentech, Inc. | Amp for use in treating microbial disorders |
US7892760B2 (en) | 2007-11-19 | 2011-02-22 | Celera Corporation | Lung cancer markers, and uses thereof |
KR20100087330A (en) | 2007-11-19 | 2010-08-04 | 제넨테크, 인크. | Compositions and methods for inhibiting tumor progression |
KR101945394B1 (en) | 2007-11-27 | 2019-02-07 | 더 유니버시티 오브 브리티쉬 콜롬비아 | 14-3-3 ETA antibodies and uses thereof for the diagnosis and treatment of arthritis |
AU2008328781A1 (en) | 2007-11-27 | 2009-06-04 | Ablynx N.V. | Amino acid sequences directed against heterodimeric cytokines and/or their receptors and polypeptides comprising the same |
TWI580694B (en) | 2007-11-30 | 2017-05-01 | 建南德克公司 | Anti-vegf antibodies |
US8426153B2 (en) | 2007-12-03 | 2013-04-23 | Carnegie Mellon University | Linked peptides fluorogenic biosensors |
DK2851374T3 (en) | 2007-12-14 | 2017-06-19 | Bristol Myers Squibb Co | Binding molecules to the human OX40 receptor |
US8557243B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | EFGR antibodies comprising modular recognition domains |
SG189769A1 (en) | 2008-01-03 | 2013-05-31 | Scripps Research Inst | Antibody targeting through a modular recognition domain |
US8557242B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | ERBB2 antibodies comprising modular recognition domains |
US8454960B2 (en) | 2008-01-03 | 2013-06-04 | The Scripps Research Institute | Multispecific antibody targeting and multivalency through modular recognition domains |
US8574577B2 (en) | 2008-01-03 | 2013-11-05 | The Scripps Research Institute | VEGF antibodies comprising modular recognition domains |
US8962803B2 (en) | 2008-02-29 | 2015-02-24 | AbbVie Deutschland GmbH & Co. KG | Antibodies against the RGM A protein and uses thereof |
CN101965362A (en) | 2008-03-05 | 2011-02-02 | 埃博灵克斯股份有限公司 | Novel antigens is in conjunction with dimer-mixture and its production and application |
EP2098536A1 (en) | 2008-03-05 | 2009-09-09 | 4-Antibody AG | Isolation and identification of antigen- or ligand-specific binding proteins |
US9873957B2 (en) | 2008-03-13 | 2018-01-23 | Dyax Corp. | Libraries of genetic packages comprising novel HC CDR3 designs |
TWI461210B (en) | 2008-03-18 | 2014-11-21 | Abbvie Inc | Methods for treating psoriasis |
EP2105742A1 (en) | 2008-03-26 | 2009-09-30 | Sanofi-Aventis | Use of cathepsin C |
AU2009228158B2 (en) | 2008-03-27 | 2014-02-27 | Zymogenetics, Inc. | Compositions and methods for inhibiting PDGFRbeta and VEGF-A |
WO2009124090A1 (en) * | 2008-03-31 | 2009-10-08 | Genentech, Inc. | Compositions and methods for treating and diagnosing asthma |
JP2011516520A (en) | 2008-04-07 | 2011-05-26 | アブリンクス エン.ヴェー. | Amino acid sequence having directivity in Notch pathway and use thereof |
AU2008201871A1 (en) * | 2008-04-16 | 2009-11-26 | Deutsches Krebsforschungszentrum Stiftung Des Oeffentlichen Rechts | Inhibition of angiogenesis and tumor metastasis |
WO2009132287A2 (en) | 2008-04-24 | 2009-10-29 | Dyax Corp. | Libraries of genetic packages comprising novel hc cdr1, cdr2, and cdr3 and novel lc cdr1, cdr2, and cdr3 designs |
MX2010011955A (en) | 2008-04-29 | 2011-01-21 | Abbott Lab | Dual variable domain immunoglobulins and uses thereof. |
SG176464A1 (en) | 2008-05-09 | 2011-12-29 | Agency Science Tech & Res | Diagnosis and treatment of kawasaki disease |
CN104558178A (en) | 2008-05-09 | 2015-04-29 | Abbvie公司 | Antibodies to receptor of advanced glycation end products (rage) and uses thereof |
EP2285833B1 (en) | 2008-05-16 | 2014-12-17 | Ablynx N.V. | AMINO ACID SEQUENCES DIRECTED AGAINST CXCR4 AND OTHER GPCRs AND COMPOUNDS COMPRISING THE SAME |
NZ589436A (en) | 2008-06-03 | 2012-12-21 | Abbott Lab | Dual variable domain immunoglobulins and uses thereof |
US9035027B2 (en) | 2008-06-03 | 2015-05-19 | Abbvie Inc. | Dual variable domain immunoglobulins and uses thereof |
PT2285408T (en) | 2008-06-05 | 2019-02-01 | Ablynx Nv | Amino acid sequences directed against envelope proteins of a virus and polypeptides comprising the same for the treatment of viral diseases |
CA2728308A1 (en) | 2008-06-20 | 2009-12-23 | Wyeth Llc | Compositions and methods of use of orf1358 from beta-hemolytic streptococcal strains |
NZ590074A (en) | 2008-07-08 | 2012-12-21 | Abbott Lab | Prostaglandin e2 dual variable domain immunoglobulins and uses thereof |
EP2310049A4 (en) | 2008-07-08 | 2013-06-26 | Abbvie Inc | Prostaglandin e2 binding proteins and uses thereof |
JP2011527572A (en) | 2008-07-09 | 2011-11-04 | バイオジェン・アイデック・エムエイ・インコーポレイテッド | Composition comprising a LINGO antibody or fragment |
KR20110036638A (en) | 2008-07-25 | 2011-04-07 | 리차드 더블유. 와그너 | Protein screening methods |
WO2010016806A1 (en) | 2008-08-08 | 2010-02-11 | Agency For Science, Technology And Research (A*Star) | Vhz for diagnosis and treatment of cancers |
US8795981B2 (en) | 2008-08-08 | 2014-08-05 | Molecular Devices, Llc | Cell detection |
MX2011001409A (en) | 2008-08-14 | 2011-03-29 | Cephalon Australia Pty Ltd | Anti-il-12/il-23 antibodies. |
BRPI0918648A2 (en) | 2008-09-03 | 2019-09-03 | Genentech Inc | multispecific antibodies |
BRPI0823049A2 (en) | 2008-09-07 | 2015-06-16 | Glyconex Inc | Anti-extended type 1 glycosphingolipid antibodies, derivatives thereof and use. |
US9075065B2 (en) | 2008-09-12 | 2015-07-07 | Dako Denmark A/S | Prostate cancer biomarker |
US8417011B2 (en) | 2008-09-18 | 2013-04-09 | Molecular Devices (New Milton) Ltd. | Colony detection |
US20100093563A1 (en) * | 2008-09-22 | 2010-04-15 | Robert Anthony Williamson | Methods and vectors for display of molecules and displayed molecules and collections |
WO2010033237A2 (en) * | 2008-09-22 | 2010-03-25 | Calmune Corporation | Methods for creating diversity in libraries and libraries, display vectors and methods, and displayed molecules |
WO2010034779A2 (en) | 2008-09-24 | 2010-04-01 | The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin | Composition and method for treatment of preterm labor |
LT2334705T (en) | 2008-09-26 | 2017-03-27 | Ucb Biopharma Sprl | Biological products |
WO2010043650A2 (en) | 2008-10-14 | 2010-04-22 | Ablynx Nv | Amino acid sequences directed against cellular receptors for viruses and bacteria |
CA2738243C (en) | 2008-10-29 | 2020-09-29 | Wyeth Llc | Formulations of single domain antigen binding molecules |
US10118962B2 (en) | 2008-10-29 | 2018-11-06 | Ablynx N.V. | Methods for purification of single domain antigen binding molecules |
CA2742968C (en) | 2008-11-07 | 2020-06-09 | Fabrus Llc | Combinatorial antibody libraries and uses thereof |
JP5823871B2 (en) | 2008-12-10 | 2015-11-25 | アブリンクス エン.ヴェー. | Amino acid sequences directed against the Angiopoietin / Tie system for the treatment of diseases and disorders associated with angiogenesis and polypeptides comprising the same |
EP2373689A1 (en) | 2008-12-12 | 2011-10-12 | MedImmune, LLC | Crystals and structure of a human igg fc variant with enhanced fcrn binding |
TW201029662A (en) | 2008-12-19 | 2010-08-16 | Glaxo Group Ltd | Novel antigen binding proteins |
AU2009329501B2 (en) | 2008-12-19 | 2015-11-26 | Ablynx N.V. | Genetic immunization for producing immunoglobulins against cell-associated antigens such as P2X7, CXCR7 or CXCR4 |
WO2010078526A1 (en) | 2008-12-31 | 2010-07-08 | Biogen Idec Ma Inc. | Anti-lymphotoxin antibodies |
JP2012515544A (en) | 2009-01-21 | 2012-07-12 | オックスフォード ビオトヘラペウトイクス エルティーディー. | PTA089 protein |
US20100260752A1 (en) | 2009-01-23 | 2010-10-14 | Biosynexus Incorporated | Opsonic and protective antibodies specific for lipoteichoic acid of gram positive bacteria |
AU2010208637A1 (en) | 2009-01-29 | 2011-08-04 | Abbvie Inc. | IL-1 binding proteins |
EP2219029A1 (en) | 2009-01-30 | 2010-08-18 | Sanofi-Aventis | Test systems, methods and uses involving AS160 protein |
ES2712732T3 (en) | 2009-02-17 | 2019-05-14 | Cornell Res Foundation Inc | Methods and kits for the diagnosis of cancer and the prediction of therapeutic value |
US9671400B2 (en) | 2009-02-19 | 2017-06-06 | Dako Denmark A/S | Conjugate molecules |
US8030026B2 (en) | 2009-02-24 | 2011-10-04 | Abbott Laboratories | Antibodies to troponin I and methods of use thereof |
WO2010100437A2 (en) | 2009-03-05 | 2010-09-10 | University Of Sheffield | Production of protein |
SG173705A1 (en) | 2009-03-05 | 2011-09-29 | Abbott Lab | Il-17 binding proteins |
WO2010102175A1 (en) | 2009-03-05 | 2010-09-10 | Medarex, Inc. | Fully human antibodies specific to cadm1 |
WO2010100135A1 (en) | 2009-03-05 | 2010-09-10 | Ablynx N.V. | Novel antigen binding dimer-complexes, methods of making/avoiding and uses thereof |
US8283162B2 (en) | 2009-03-10 | 2012-10-09 | Abbott Laboratories | Antibodies relating to PIVKAII and uses thereof |
SI3260136T1 (en) | 2009-03-17 | 2021-05-31 | Theraclone Sciences, Inc. | Human immunodeficiency virus (hiv) -neutralizing antibodies |
GB0905023D0 (en) | 2009-03-24 | 2009-05-06 | Univ Erasmus Medical Ct | Binding molecules |
JP5616428B2 (en) | 2009-04-07 | 2014-10-29 | ロシュ グリクアート アクチェンゲゼルシャフト | Trivalent bispecific antibody |
CN105399828B (en) | 2009-04-10 | 2021-01-15 | 埃博灵克斯股份有限公司 | Improved amino acid sequences directed against IL-6R and polypeptides comprising the same for the treatment of IL-6R related diseases and disorders |
AP2011005984A0 (en) | 2009-04-20 | 2011-12-31 | Oxford Biotherapeutics Ltd | Antibodies specific to cadherin-17. |
CA2759506A1 (en) * | 2009-04-23 | 2010-10-28 | Theraclone Sciences, Inc. | Granulocyte-macrophage colony-stimulating factor (gm-csf) neutralizing antibodies |
CA2759370C (en) | 2009-04-30 | 2020-02-11 | Peter Schotte | Method for the production of domain antibodies |
AU2010249470B2 (en) * | 2009-05-20 | 2015-06-25 | Novimmune S.A. | Synthetic Polypeptide Libraries And Methods For Generating Naturally Diversified Polypeptide Variants |
DK2435568T3 (en) | 2009-05-29 | 2014-09-08 | Morphosys Ag | Collection of synthetic antibodies to treat disease |
EP2438087B1 (en) | 2009-06-05 | 2017-05-10 | Ablynx N.V. | Trivalent anti human respiratory syncytial virus (hrsv) nanobody constructs for the prevention and/or treatment of respiratory tract infections |
US9676845B2 (en) | 2009-06-16 | 2017-06-13 | Hoffmann-La Roche, Inc. | Bispecific antigen binding proteins |
JP5683581B2 (en) * | 2009-06-30 | 2015-03-11 | リサーチ ディベロップメント ファウンデーション | Immunoglobulin Fc polypeptide |
WO2011000054A1 (en) | 2009-07-03 | 2011-01-06 | Avipep Pty Ltd | Immuno-conjugates and methods for producing them |
IE20090514A1 (en) | 2009-07-06 | 2011-02-16 | Opsona Therapeutics Ltd | Humanised antibodies and uses therof |
US9150640B2 (en) | 2009-07-10 | 2015-10-06 | Ablynx N.V. | Method for the production of variable domains |
TW201106972A (en) | 2009-07-27 | 2011-03-01 | Genentech Inc | Combination treatments |
CA2771575A1 (en) | 2009-08-29 | 2011-03-03 | Abbott Laboratories | Therapeutic dll4 binding proteins |
EP2293072A1 (en) | 2009-08-31 | 2011-03-09 | Sanofi-Aventis | Use of cathepsin H |
JP5715137B2 (en) | 2009-08-31 | 2015-05-07 | アボット・ラボラトリーズAbbott Laboratories | Biomarkers and their use for prediction of major adverse cardiac events |
CA2772628A1 (en) | 2009-09-01 | 2011-03-10 | Abbott Laboratories | Dual variable domain immunoglobulins and uses thereof |
CA2772715C (en) | 2009-09-02 | 2019-03-26 | Genentech, Inc. | Mutant smoothened and methods of using the same |
EP2473528B1 (en) | 2009-09-03 | 2014-12-03 | Ablynx N.V. | Stable formulations of polypeptides and uses thereof |
ES2788869T3 (en) | 2009-09-03 | 2020-10-23 | Merck Sharp & Dohme | Anti-GITR antibodies |
EP2477654A4 (en) | 2009-09-14 | 2013-01-23 | Abbott Lab | Methods for treating psoriasis |
JP2013504602A (en) * | 2009-09-14 | 2013-02-07 | ダイアックス コーポレーション | Newly designed gene package library containing HCCR3 |
SG10201408401RA (en) | 2009-09-16 | 2015-01-29 | Genentech Inc | Coiled coil and/or tether containing protein complexes and uses thereof |
US20110189183A1 (en) | 2009-09-18 | 2011-08-04 | Robert Anthony Williamson | Antibodies against candida, collections thereof and methods of use |
GB201005063D0 (en) | 2010-03-25 | 2010-05-12 | Ucb Pharma Sa | Biological products |
US8568726B2 (en) | 2009-10-06 | 2013-10-29 | Medimmune Limited | RSV specific binding molecule |
US8518405B2 (en) | 2009-10-08 | 2013-08-27 | The University Of North Carolina At Charlotte | Tumor specific antibodies and uses therefor |
TW201117824A (en) | 2009-10-12 | 2011-06-01 | Amgen Inc | Use of IL-17 receptor a antigen binding proteins |
EP2470569A1 (en) | 2009-10-13 | 2012-07-04 | Oxford Biotherapeutics Ltd. | Antibodies against epha10 |
AR078651A1 (en) | 2009-10-15 | 2011-11-23 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
WO2011046457A1 (en) | 2009-10-16 | 2011-04-21 | Auckland Uniservices Limited | Anti-neoplastic uses of artemin antagonists |
WO2011047680A1 (en) | 2009-10-20 | 2011-04-28 | Dako Denmark A/S | Immunochemical detection of single target entities |
UY32979A (en) | 2009-10-28 | 2011-02-28 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
JO3437B1 (en) | 2009-10-30 | 2019-10-20 | Esai R & D Man Co Ltd | Improved anti human Fraktalkine antibodies and uses thereof |
TW201121568A (en) | 2009-10-31 | 2011-07-01 | Abbott Lab | Antibodies to receptor for advanced glycation end products (RAGE) and uses thereof |
EP2496605A1 (en) | 2009-11-02 | 2012-09-12 | Oxford Biotherapeutics Ltd. | Ror1 as therapeutic and diagnostic target |
NZ599761A (en) | 2009-11-04 | 2014-04-30 | Merck Sharp & Dohme | Engineered anti-tslp antibody |
JP6007420B2 (en) | 2009-11-04 | 2016-10-12 | ファブラス エルエルシー | Antibody optimization method based on affinity maturation |
US20110165648A1 (en) | 2009-11-04 | 2011-07-07 | Menno Van Lookeren Campagne | Co-crystal structure of factor D and anti-factor D antibody |
EP2496944A2 (en) | 2009-11-05 | 2012-09-12 | Novartis AG | Biomarkers predictive of progression of fibrosis |
WO2011058087A1 (en) | 2009-11-11 | 2011-05-19 | Gentian As | Immunoassay for assessing related analytes of different origin |
US9644022B2 (en) | 2009-11-30 | 2017-05-09 | Ablynx N.V. | Amino acid sequences directed against human respiratory syncytial virus (HRSV) and polypeptides comprising the same for the prevention and/or treatment of respiratory tract infections |
EP3135302A1 (en) | 2009-12-02 | 2017-03-01 | Imaginab, Inc. | J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use |
CN102656190A (en) | 2009-12-08 | 2012-09-05 | 雅培股份有限两合公司 | Monoclonal antibodies against the RGM A protein for use in the treatment of retinal nerve fiber layer degeneration |
US8486397B2 (en) | 2009-12-11 | 2013-07-16 | Genentech, Inc. | Anti-VEGF-C antibodies and methods using same |
US8962807B2 (en) | 2009-12-14 | 2015-02-24 | Ablynx N.V. | Single variable domain antibodies against OX40L, constructs and therapeutic use |
PL2515941T3 (en) | 2009-12-21 | 2020-04-30 | F. Hoffmann-La Roche Ag | Pharmaceutical formulation of bevacizumab |
AU2010336485B2 (en) | 2009-12-23 | 2015-03-26 | Genentech, Inc. | Anti-Bv8 antibodies and uses thereof |
WO2011075786A1 (en) | 2009-12-23 | 2011-06-30 | Avipep Pty Ltd | Immuno-conjugates and methods for producing them 2 |
WO2011083141A2 (en) | 2010-01-08 | 2011-07-14 | Ablynx Nv | Method for generation of immunoglobulin sequences by using lipoprotein particles |
TWI513466B (en) | 2010-01-20 | 2015-12-21 | Boehringer Ingelheim Int | Anticoagulant antidotes |
EP2528947A4 (en) | 2010-01-28 | 2013-09-18 | Glaxo Group Ltd | Cd127 binding proteins |
EP2354159A1 (en) | 2010-02-05 | 2011-08-10 | RWTH Aachen | CCL17 inhibitors for use in T helper cell-driven diseases |
AU2011214465A1 (en) | 2010-02-10 | 2012-08-30 | Novartis Ag | Methods and compounds for muscle growth |
US9120855B2 (en) | 2010-02-10 | 2015-09-01 | Novartis Ag | Biologic compounds directed against death receptor 5 |
CN102753148B (en) | 2010-02-11 | 2018-01-26 | 埃博灵克斯股份有限公司 | For preparing the method and composition of aerosol |
ES2519348T3 (en) | 2010-02-18 | 2014-11-06 | Genentech, Inc. | Neurregulin antagonists and their use in cancer treatment |
PE20130580A1 (en) | 2010-03-02 | 2013-06-02 | Abbvie Inc | THERAPEUTIC BINDING PROTEINS TO DLL4 |
UA108227C2 (en) | 2010-03-03 | 2015-04-10 | ANTIGENCY PROTEIN | |
GB201003701D0 (en) | 2010-03-05 | 2010-04-21 | Cilian Ag | System for the expression of a protein |
SG10201504808XA (en) | 2010-03-17 | 2015-07-30 | Abbott Res Bv | Anti-Nerve Growth Factor (NGF) Antibody Compositions |
BR112012022044A2 (en) | 2010-03-24 | 2020-08-25 | Genentech Inc | ''antibody, immunoconjugate, pharmaceutical formulation, antibody use, treatment method, isolated bispecific antibody and host cell''. |
US8937164B2 (en) | 2010-03-26 | 2015-01-20 | Ablynx N.V. | Biological materials related to CXCR7 |
TW201138821A (en) | 2010-03-26 | 2011-11-16 | Roche Glycart Ag | Bispecific antibodies |
MX336196B (en) | 2010-04-15 | 2016-01-11 | Abbvie Inc | Amyloid-beta binding proteins. |
SG185415A1 (en) | 2010-05-06 | 2012-12-28 | Novartis Ag | Compositions and methods of use for therapeutic low density lipoprotein - related protein 6 (lrp6) multivalent antibodies |
PE20130207A1 (en) | 2010-05-06 | 2013-02-28 | Novartis Ag | ANTIBODIES ANTAGONISTS TO LRP6 (LOW DENSITY LIPOPROTEIN-RELATED PROTEIN 6) AND COMPOSITIONS |
AU2011252883B2 (en) | 2010-05-14 | 2015-09-10 | Abbvie Inc. | IL-1 binding proteins |
WO2011144749A1 (en) | 2010-05-20 | 2011-11-24 | Ablynx Nv | Biological materials related to her3 |
EP2577309B1 (en) | 2010-05-25 | 2016-11-23 | Carnegie Mellon University | Targeted probes of cellular physiology |
WO2011147834A1 (en) | 2010-05-26 | 2011-12-01 | Roche Glycart Ag | Antibodies against cd19 and uses thereof |
AR081556A1 (en) | 2010-06-03 | 2012-10-03 | Glaxo Group Ltd | HUMANIZED ANTIGEN UNION PROTEINS |
NZ602840A (en) | 2010-06-03 | 2014-11-28 | Genentech Inc | Immuno-pet imaging of antibodies and immunoconjugates and uses therefor |
WO2011161545A2 (en) | 2010-06-04 | 2011-12-29 | The Netherlands Cancer Institute | Non-hydrolyzable protein conjugates, methods and compositions related thereto |
WO2011158019A1 (en) | 2010-06-16 | 2011-12-22 | Adjuvantix Limited | Polypeptide vaccine |
CN103080136B (en) | 2010-06-18 | 2015-08-12 | 霍夫曼-拉罗奇有限公司 | Anti-Axl antibody and using method |
WO2011161119A1 (en) | 2010-06-22 | 2011-12-29 | F. Hoffmann-La Roche Ag | Antibodies against insulin-like growth factor i receptor and uses thereof |
WO2011161189A1 (en) | 2010-06-24 | 2011-12-29 | F. Hoffmann-La Roche Ag | Anti-hepsin antibodies and methods of use |
WO2011161263A1 (en) | 2010-06-25 | 2011-12-29 | Ablynx Nv | Pharmaceutical compositions for cutaneous administration |
US20120009196A1 (en) | 2010-07-08 | 2012-01-12 | Abbott Laboratories | Monoclonal antibodies against hepatitis c virus core protein |
NZ605400A (en) | 2010-07-09 | 2015-05-29 | Biogen Idec Hemophilia Inc | Chimeric clotting factors |
UY33492A (en) | 2010-07-09 | 2012-01-31 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
BR112013000340A2 (en) | 2010-07-09 | 2016-05-31 | Genentech Inc | isolated antibody that binds neuropillin-1 (nrp1), isolated nucleic acid, host cell, method of producing an antibody, immunoconjugate and method of detecting nrp1 in a biological sample |
US20120100166A1 (en) | 2010-07-15 | 2012-04-26 | Zyngenia, Inc. | Ang-2 Binding Complexes and Uses Thereof |
CA3086837C (en) | 2010-07-16 | 2023-03-07 | Adimab, Llc | Libraries comprising segmental pools, and methods for their preparation and use |
AU2011282476B2 (en) | 2010-07-20 | 2015-08-20 | Cephalon Australia Pty Ltd | Anti-IL-23 heterodimer specific antibodies |
WO2012010582A1 (en) | 2010-07-21 | 2012-01-26 | Roche Glycart Ag | Anti-cxcr5 antibodies and methods of use |
US9120862B2 (en) | 2010-07-26 | 2015-09-01 | Abbott Laboratories | Antibodies relating to PIVKA-II and uses thereof |
AU2011285852B2 (en) | 2010-08-03 | 2014-12-11 | Abbvie Inc. | Dual variable domain immunoglobulins and uses thereof |
EP2600895A1 (en) | 2010-08-03 | 2013-06-12 | Hoffmann-La Roche AG | Chronic lymphocytic leukemia (cll) biomarkers |
RU2013106217A (en) | 2010-08-05 | 2014-09-10 | Ф. Хоффманн-Ля Рош Аг | HYBRID PROTEIN FROM ANTIBODIES AGAINST MHC AND ANTIVIRAL CYTOKINE |
EP2603526A1 (en) | 2010-08-13 | 2013-06-19 | Medimmune Limited | Monomeric polypeptides comprising variant fc regions and methods of use |
EA037977B1 (en) | 2010-08-13 | 2021-06-18 | Роше Гликарт Аг | Anti-fap antibodies, methods of production and use thereof |
WO2012020038A1 (en) | 2010-08-13 | 2012-02-16 | Roche Glycart Ag | Anti-tenascin-c a2 antibodies and methods of use |
WO2012024187A1 (en) | 2010-08-14 | 2012-02-23 | Abbott Laboratories | Amyloid-beta binding proteins |
WO2012022734A2 (en) | 2010-08-16 | 2012-02-23 | Medimmune Limited | Anti-icam-1 antibodies and methods of use |
HUE058226T2 (en) | 2010-08-19 | 2022-07-28 | Zoetis Belgium S A | Anti-ngf antibodies and their use |
MY162825A (en) | 2010-08-20 | 2017-07-31 | Novartis Ag | Antibodies for epidermal growth factor receptor 3 (her3) |
WO2012025530A1 (en) | 2010-08-24 | 2012-03-01 | F. Hoffmann-La Roche Ag | Bispecific antibodies comprising a disulfide stabilized - fv fragment |
TW201215405A (en) | 2010-08-25 | 2012-04-16 | Hoffmann La Roche | Antibodies against IL-18R1 and uses thereof |
KR20130139884A (en) | 2010-08-26 | 2013-12-23 | 애브비 인코포레이티드 | Dual variable domain immunoglobulins and uses thereof |
ES2920140T3 (en) | 2010-08-31 | 2022-08-01 | Theraclone Sciences Inc | Human immunodeficiency virus (HIV) neutralizing antibodies |
RU2013114360A (en) | 2010-08-31 | 2014-10-10 | Дженентек, Инк. | BIOMARKERS AND TREATMENT METHODS |
KR20130096731A (en) | 2010-09-08 | 2013-08-30 | 할로자임, 아이엔씨 | Methods for assessing and identifying or evolving conditionally active therapeutic proteins |
KR101527297B1 (en) | 2010-09-09 | 2015-06-26 | 화이자 인코포레이티드 | 4-1bb binding molecules |
WO2012038744A2 (en) | 2010-09-22 | 2012-03-29 | Genome Research Limited | Detecting mutations |
GB201016494D0 (en) | 2010-09-30 | 2010-11-17 | Queen Mary Innovation Ltd | Polypeptide |
US8497138B2 (en) | 2010-09-30 | 2013-07-30 | Genetix Limited | Method for cell selection |
US8481680B2 (en) | 2010-10-05 | 2013-07-09 | Genentech, Inc. | Mutant smoothened and methods of using the same |
ES2660895T3 (en) | 2010-10-29 | 2018-03-26 | Ablynx N.V. | Method for the production of individual variable domains of immunoglobulin |
MX352929B (en) | 2010-11-05 | 2017-12-13 | Zymeworks Inc | Stable heterodimeric antibody design with mutations in the fc domain. |
CN103201627B (en) | 2010-11-08 | 2016-10-12 | 丹麦达科有限公司 | In histological sample, single target molecules is quantitative |
TWI619730B (en) | 2010-11-08 | 2018-04-01 | 諾華公司 | Chemokine receptor binding polypeptides |
TW201300417A (en) | 2010-11-10 | 2013-01-01 | Genentech Inc | Methods and compositions for neural disease immunotherapy |
JP6253986B2 (en) | 2010-11-19 | 2017-12-27 | モルフォシス・アーゲー | Collection and its usage |
CA2817161C (en) | 2010-12-06 | 2019-04-02 | Dako Denmark A/S | Combined histological stain |
BR122020012255B1 (en) | 2010-12-16 | 2022-08-09 | Genentech, Inc | USE OF AN ANTI-IL-13 ANTIBODY, USES OF A TH2 PATHWAY INHIBITOR AND ANTI-PERIOSTIN ANTIBODIES |
TW201238974A (en) | 2010-12-17 | 2012-10-01 | Sanofi Sa | MiRNAs in joint disease |
TW201241179A (en) | 2010-12-17 | 2012-10-16 | Sanofi Sa | MiRNAs in joint disease |
TW201239097A (en) | 2010-12-17 | 2012-10-01 | Sanofi Sa | MiRNAs in joint disease |
UY33807A (en) | 2010-12-17 | 2012-07-31 | Sanofi Sa | miRNAs as indicators of tissue status or of diseases such as osteoarthritis |
AU2011349443B2 (en) | 2010-12-20 | 2015-12-24 | Genentech, Inc. | Anti-mesothelin antibodies and immunoconjugates |
EP2655607A4 (en) | 2010-12-21 | 2014-05-14 | Univ North Carolina | Methods and compositions for producing active vitamin k-dependent proteins |
TW201307388A (en) | 2010-12-21 | 2013-02-16 | Abbott Lab | IL-1 binding proteins |
RU2627171C2 (en) | 2010-12-21 | 2017-08-03 | Эббви Инк. | Il-1 alpha and beta bispecific immunoglobulins with double variable domains and their application |
KR20130118925A (en) | 2010-12-22 | 2013-10-30 | 제넨테크, 인크. | Anti-pcsk9 antibodies and methods of use |
CN103384831B (en) | 2010-12-23 | 2016-02-10 | 霍夫曼-拉罗奇有限公司 | Polypeptide dimer is detected by bivalent binders |
ES2605493T3 (en) | 2010-12-23 | 2017-03-14 | F. Hoffmann-La Roche Ag | Detection of a post-translationally modified polypeptide by a bivalent binding agent |
WO2012085111A1 (en) | 2010-12-23 | 2012-06-28 | F. Hoffmann-La Roche Ag | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
CA2817448C (en) | 2010-12-23 | 2019-01-22 | F. Hoffmann-La Roche Ag | Binding agent |
US20140038842A1 (en) | 2010-12-28 | 2014-02-06 | Xoma Technology | Cell surface display using pdz domains |
EP2471554A1 (en) | 2010-12-28 | 2012-07-04 | Hexal AG | Pharmaceutical formulation comprising a biopharmaceutical drug |
WO2012109133A1 (en) | 2011-02-07 | 2012-08-16 | Research Development Foundation | Engineered immunoglobulin fc polypeptides |
AU2012215572A1 (en) | 2011-02-10 | 2013-05-02 | Roche Glycart Ag | Improved immunotherapy |
WO2012109624A2 (en) | 2011-02-11 | 2012-08-16 | Zyngenia, Inc. | Monovalent and multivalent multispecific complexes and uses thereof |
AR085403A1 (en) | 2011-02-28 | 2013-09-25 | Hoffmann La Roche | MONOVALENT PROTEINS THAT JOIN ANTIGENS |
RU2607038C2 (en) | 2011-02-28 | 2017-01-10 | Ф. Хоффманн-Ля Рош Аг | Antigen-binding proteins |
EP2686016B1 (en) | 2011-03-14 | 2019-05-01 | Cellmid Limited | Antibody recognizing n-domain of midkine |
AR085911A1 (en) | 2011-03-16 | 2013-11-06 | Sanofi Sa | SAFE THERAPEUTIC DOSE OF A SIMILAR PROTEIN TO AN ANTIBODY WITH VUAL REGION |
AU2012234284B2 (en) | 2011-03-28 | 2015-10-08 | Ablynx Nv | Bispecific anti-CXCR7 immunoglobulin single variable domains |
ES2688591T3 (en) | 2011-03-28 | 2018-11-05 | Ablynx N.V. | Method for producing solid formulations comprising individual variable domains of immunoglobulin |
MY163539A (en) | 2011-03-29 | 2017-09-15 | Roche Glycart Ag | Antibody fc variants |
MA34978B1 (en) | 2011-03-30 | 2014-03-01 | Boehringer Ingelheim Int | ANTIDOTES FOR ANTICOAGULANTS |
MX342240B (en) | 2011-04-07 | 2016-09-21 | Genentech Inc | Anti-fgfr4 antibodies and methods of use. |
EP2511293A1 (en) | 2011-04-13 | 2012-10-17 | LEK Pharmaceuticals d.d. | A method for controlling the main complex N-glycan structures and the acidic variants and variability in bioprocesses producing recombinant proteins |
CA2832907C (en) | 2011-04-19 | 2020-07-14 | Dako Denmark A/S | New method for enzyme-mediated signal amplification |
KR20140031217A (en) | 2011-04-20 | 2014-03-12 | 로슈 글리카트 아게 | Method and constructs for the ph dependent passage of the blood-brain-barrier |
AU2012245073B2 (en) | 2011-04-21 | 2016-02-11 | Garvan Institute Of Medical Research | Modified variable domain molecules and methods for producing and using them b |
EP2518157A1 (en) | 2011-04-26 | 2012-10-31 | Sanofi | Test Systems and methods for identifying a compound altering cellular DDR activity |
WO2012149197A2 (en) | 2011-04-27 | 2012-11-01 | Abbott Laboratories | Methods for controlling the galactosylation profile of recombinantly-expressed proteins |
EA201892619A1 (en) | 2011-04-29 | 2019-04-30 | Роше Гликарт Аг | IMMUNOCONJUGATES CONTAINING INTERLEUKIN-2 MUTANT POLYPETIPS |
UA117218C2 (en) | 2011-05-05 | 2018-07-10 | Мерк Патент Гмбх | Amino acid sequences directed against il-17a, il-17f and/or il17-a/f and polypeptides comprising the same |
BR112013028655B1 (en) | 2011-05-06 | 2022-08-16 | Zoetis Services Llc | ANTI-NEURONAL GROWTH FACTOR ANTIBODIES, PHARMACEUTICAL COMPOSITION COMPRISING THEM AND KIT FOR THE TREATMENT OF FELINE PAIN |
CA2835094C (en) | 2011-05-06 | 2020-12-22 | David Gearing | Anti-nerve growth factor antibodies and methods of preparing and using the same |
PL3498732T3 (en) | 2011-05-06 | 2022-02-28 | Zoetis Services Llc | Anti-nerve growth factor antibodies and methods of preparing and using the same |
ES2704007T3 (en) | 2011-05-06 | 2019-03-13 | Nexvet Australia Pty Ltd | Anti-nerve growth factor antibodies and procedures for preparing and using them |
GB201114858D0 (en) | 2011-08-29 | 2011-10-12 | Nvip Pty Ltd | Anti-nerve growth factor antibodies and methods of using the same |
US9534039B2 (en) | 2011-05-09 | 2017-01-03 | Ablynx N.V. | Method for the production of immunoglobulin single variable domains |
WO2012155019A1 (en) | 2011-05-12 | 2012-11-15 | Genentech, Inc. | Multiple reaction monitoring lc-ms/ms method to detect therapeutic antibodies in animal samples using framework signature pepides |
SG194917A1 (en) | 2011-05-16 | 2013-12-30 | Genentech Inc | Fgfr1 agonists and methods of use |
CN106432484B (en) | 2011-05-17 | 2020-10-30 | 洛克菲勒大学 | Neutralizing antibodies to human immunodeficiency virus and methods of use thereof |
EP2714738B1 (en) | 2011-05-24 | 2018-10-10 | Zyngenia, Inc. | Multivalent and monovalent multispecific complexes and their uses |
EP2714736A1 (en) | 2011-05-27 | 2014-04-09 | Ablynx N.V. | Inhibition of bone resorption with rankl binding peptides |
US9580480B2 (en) | 2011-05-31 | 2017-02-28 | Massachusetts Institute Of Technology | Cell-directed synthesis of multifunctional nanopatterns and nanomaterials |
HUE038509T2 (en) | 2011-06-10 | 2018-10-29 | Medimmune Ltd | Anti-pseudomonas psl binding molecules and uses thereof |
MX343580B (en) | 2011-06-13 | 2016-11-10 | Csl Ltd | Antibodies against g-csfr and uses thereof. |
US8623666B2 (en) | 2011-06-15 | 2014-01-07 | Hoffmann-La Roche Inc. | Method for detecting erythropoietin (EPO) receptor using anti-human EPO receptor antibodies |
EP2537532A1 (en) | 2011-06-22 | 2012-12-26 | J. Stefan Institute | Cathepsin-binding compounds bound to a nanodevice and their diagnostic and therapeutic use |
SI2723769T2 (en) | 2011-06-23 | 2022-09-30 | Ablynx Nv | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains |
EP2723772A1 (en) | 2011-06-23 | 2014-04-30 | Ablynx N.V. | Immunoglobulin single variable domains directed against ige |
KR20240033183A (en) | 2011-06-23 | 2024-03-12 | 아블린쓰 엔.브이. | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains |
EP4350345A2 (en) | 2011-06-23 | 2024-04-10 | Ablynx N.V. | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobin single variable domains |
EA031181B1 (en) | 2011-06-28 | 2018-11-30 | Оксфорд Байотерепьютикс Лтд. | Antibody or an antigen binding fragment thereof that specifically binds to bst1, nucleic acids encoding chains thereof, host cell and method provided for making this antibody and use thereof for treating cancer and inflammatory diseases |
LT2726094T (en) | 2011-06-28 | 2017-02-10 | Oxford Biotherapeutics Ltd | Therapeutic and diagnostic target |
BR112013030472A2 (en) | 2011-06-30 | 2019-09-24 | Genentech Inc | pharmaceutical formulation, article of manufacture and method |
WO2013009521A2 (en) | 2011-07-13 | 2013-01-17 | Abbvie Inc. | Methods and compositions for treating asthma using anti-il-13 antibodies |
WO2013012733A1 (en) | 2011-07-15 | 2013-01-24 | Biogen Idec Ma Inc. | Heterodimeric fc regions, binding molecules comprising same, and methods relating thereto |
US20140234330A1 (en) | 2011-07-22 | 2014-08-21 | Amgen Inc. | Il-17 receptor a is required for il-17c biology |
KR20140057326A (en) | 2011-08-17 | 2014-05-12 | 제넨테크, 인크. | Neuregulin antibodies and uses thereof |
WO2013026832A1 (en) | 2011-08-23 | 2013-02-28 | Roche Glycart Ag | Anti-mcsp antibodies |
SI2748202T1 (en) | 2011-08-23 | 2018-10-30 | Roche Glycart Ag | Bispecific antigen binding molecules |
US20130058936A1 (en) | 2011-08-23 | 2013-03-07 | Peter Bruenker | Bispecific antibodies specific for t-cell activating antigens and a tumor antigen and methods of use |
US20130078250A1 (en) | 2011-08-23 | 2013-03-28 | Oliver Ast | Bispecific t cell activating antigen binding molecules |
MY169358A (en) | 2011-08-23 | 2019-03-26 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
SG11201400222RA (en) | 2011-08-30 | 2014-03-28 | Nvip Pty Ltd | Caninised tumour necrosis factor antibodies and methods of using the same |
WO2013033069A1 (en) | 2011-08-30 | 2013-03-07 | Theraclone Sciences, Inc. | Human rhinovirus (hrv) antibodies |
JP6216317B2 (en) | 2011-09-09 | 2017-10-18 | メディミューン リミテッド | Anti-Siglec-15 antibody and use thereof |
GB2511928B (en) | 2011-09-14 | 2015-04-08 | Abeterno Ltd | Intracellular cell selection |
WO2013040433A1 (en) | 2011-09-15 | 2013-03-21 | Genentech, Inc. | Methods of promoting differentiation |
KR20210099167A (en) | 2011-09-19 | 2021-08-11 | 악손 뉴로사이언스 에스이 | Protein-based therapy and diagnosis of tau-mediated pathology in alzheimer's disease |
AU2012312515A1 (en) | 2011-09-19 | 2014-03-13 | Genentech, Inc. | Combination treatments comprising c-met antagonists and B-raf antagonists |
GB201116116D0 (en) | 2011-09-19 | 2011-11-02 | Univ York | Cell differentiation |
ES2806146T3 (en) | 2011-09-22 | 2021-02-16 | Amgen Inc | CD27L antigen-binding proteins |
US10138302B2 (en) | 2011-09-23 | 2018-11-27 | Ablynx N.V. | Methods for treating rheumatoid arthritis by administering interleukin-6 receptor antibodies |
GB201116702D0 (en) | 2011-09-28 | 2011-11-09 | Procure Therapeutics Ltd | Cell surface markers |
EP3495389A1 (en) | 2011-09-30 | 2019-06-12 | Teva Pharmaceuticals Australia Pty Ltd | Antibodies against tl1a and uses thereof |
US20130089562A1 (en) | 2011-10-05 | 2013-04-11 | Genenthech, Inc. | Methods of treating liver conditions using notch2 antagonists |
US9575073B2 (en) | 2011-10-10 | 2017-02-21 | Rutgers, The State University Of New Jersey | Detection of high-risk intraductal papillary mucinous neoplasm and pancreatic adenocarcinoma |
ES2687951T3 (en) | 2011-10-14 | 2018-10-30 | F. Hoffmann-La Roche Ag | Anti-HtrA1 antibodies and procedures for use |
MX2014004426A (en) | 2011-10-15 | 2014-07-09 | Genentech Inc | Scd1 antagonists for treating cancer. |
WO2013059531A1 (en) | 2011-10-20 | 2013-04-25 | Genentech, Inc. | Anti-gcgr antibodies and uses thereof |
RS20140202A1 (en) | 2011-10-24 | 2014-10-31 | Abbvie Inc. | Biospecific immunibinders directed against tnf and il-17 |
US8999331B2 (en) | 2011-10-24 | 2015-04-07 | Abbvie Inc. | Immunobinders directed against sclerostin |
CN104093739A (en) | 2011-10-24 | 2014-10-08 | 艾伯维公司 | Immunobinders directed against TNF |
GB201118359D0 (en) | 2011-10-25 | 2011-12-07 | Univ Sheffield | Pulmonary hypertension |
NZ756727A (en) | 2011-10-28 | 2022-12-23 | Teva Pharmaceuticals Australia Pty Ltd | Polypeptide constructs and uses thereof |
MX2014004991A (en) | 2011-10-28 | 2014-05-22 | Genentech Inc | Therapeutic combinations and methods of treating melanoma. |
US9265817B2 (en) | 2011-10-28 | 2016-02-23 | Patrys Limited | PAT-LM1 epitopes and methods for using same |
GB201118840D0 (en) | 2011-11-01 | 2011-12-14 | Univ Sheffield | Pulmonary hypertension II |
AU2012332263A1 (en) | 2011-11-04 | 2014-05-22 | Novartis Ag | Low density lipoprotein-related protein 6 (LRP6) - half life extender constructs |
RU2675319C2 (en) | 2011-11-04 | 2018-12-18 | Займворкс Инк. | STABLE HETERODIMERIC ANTIBODY DESIGN WITH MUTATIONS IN Fc DOMAIN |
HUE037142T2 (en) | 2011-11-11 | 2018-08-28 | Ucb Biopharma Sprl | Albumin binding antibodies and binding fragments thereof |
WO2013078170A1 (en) | 2011-11-21 | 2013-05-30 | Genentech, Inc. | Purification of anti-c-met antibodies |
BR112014012882A2 (en) | 2011-11-29 | 2017-06-13 | Genentech Inc | method, antibody, polynucleotide, host cell, hybridoma cell line, antibody use and kit |
AU2012346861A1 (en) | 2011-11-30 | 2014-06-19 | AbbVie Deutschland GmbH & Co. KG | Methods and compositions for determining responsiveness to treatment with a tnf-alpha inhibitor |
BR112014013568A8 (en) | 2011-12-05 | 2017-06-13 | Novartis Ag | epidermal growth factor 3 (her3) receptor antibodies directed to her3 domain ii |
AU2012349735B2 (en) | 2011-12-05 | 2016-05-19 | Novartis Ag | Antibodies for epidermal growth factor receptor 3 (HER3) |
US20140335084A1 (en) | 2011-12-06 | 2014-11-13 | Hoffmann-La Roche Inc. | Antibody formulation |
RU2628703C2 (en) | 2011-12-22 | 2017-08-21 | Ф. Хоффманн-Ля Рош Аг | Expression vector structure, new methods for obtaining producer cells and their application for recombinant obtaining of polypeptides |
EP2794651B1 (en) | 2011-12-22 | 2022-09-21 | F. Hoffmann-La Roche AG | Expression vector element combinations, novel production cell generation methods and their use for the recombinant production of polypeptides |
WO2013096791A1 (en) | 2011-12-23 | 2013-06-27 | Genentech, Inc. | Process for making high concentration protein formulations |
TW201333035A (en) | 2011-12-30 | 2013-08-16 | Abbvie Inc | Dual specific binding proteins directed against IL-13 and/or IL-17 |
JP6684490B2 (en) | 2012-01-09 | 2020-04-22 | ザ・スクリップス・リサーチ・インスティテュート | Ultralong complementarity determining regions and uses thereof |
US20150011431A1 (en) | 2012-01-09 | 2015-01-08 | The Scripps Research Institute | Humanized antibodies |
SI2802606T1 (en) | 2012-01-10 | 2018-08-31 | Biogen Ma Inc. | Enhancement of transport of therapeutic molecules across the blood brain barrier |
SG11201404198TA (en) | 2012-01-18 | 2014-08-28 | Genentech Inc | Anti-lrp5 antibodies and methods of use |
KR20140119114A (en) | 2012-01-18 | 2014-10-08 | 제넨테크, 인크. | Methods of using fgf19 modulators |
SG10202006762PA (en) | 2012-01-27 | 2020-08-28 | Abbvie Deutschland | Composition and method for diagnosis and treatment of diseases associated with neurite degeneration |
WO2013113641A1 (en) | 2012-01-31 | 2013-08-08 | Roche Glycart Ag | Use of nkp46 as a predictive biomarker for cancer treatment with adcc- enhanced antibodies |
CA2861124A1 (en) | 2012-02-10 | 2013-08-15 | Genentech, Inc. | Single-chain antibodies and other heteromultimers |
US20130209473A1 (en) | 2012-02-11 | 2013-08-15 | Genentech, Inc. | R-spondin translocations and methods using the same |
BR112014018005B1 (en) | 2012-02-15 | 2021-06-29 | F. Hoffmann-La Roche Ag | USE OF A NON-COVALENT IMMOBILIZED COMPLEX |
GB201203071D0 (en) | 2012-02-22 | 2012-04-04 | Ucb Pharma Sa | Biological products |
GB201203587D0 (en) | 2012-03-01 | 2012-04-11 | Univ Warwick | Modified bacterial cell |
RU2014138586A (en) | 2012-03-02 | 2016-04-20 | Рош Гликарт Аг | PROGNOSTIC BIOMARKER FOR TREATING CANCER WITH ANTIBODIES WITH AN INCREASED ANTIBODY-DEPENDENT CELL CYTOTOXICITY |
US9592289B2 (en) | 2012-03-26 | 2017-03-14 | Sanofi | Stable IgG4 based binding agent formulations |
SG11201406079TA (en) | 2012-03-27 | 2014-10-30 | Genentech Inc | Diagnosis and treatments relating to her3 inhibitors |
AU2013240090B2 (en) | 2012-03-27 | 2017-01-05 | Ventana Medical Systems, Inc. | Signaling conjugates and methods of use |
AR090549A1 (en) | 2012-03-30 | 2014-11-19 | Genentech Inc | ANTI-LGR5 AND IMMUNOCATE PLAYERS |
US9150645B2 (en) | 2012-04-20 | 2015-10-06 | Abbvie, Inc. | Cell culture methods to reduce acidic species |
US9067990B2 (en) | 2013-03-14 | 2015-06-30 | Abbvie, Inc. | Protein purification using displacement chromatography |
US9181572B2 (en) | 2012-04-20 | 2015-11-10 | Abbvie, Inc. | Methods to modulate lysine variant distribution |
KR20150006000A (en) | 2012-05-01 | 2015-01-15 | 제넨테크, 인크. | Anti-pmel17 antibodies and immunoconjugates |
US9328174B2 (en) | 2012-05-09 | 2016-05-03 | Novartis Ag | Chemokine receptor binding polypeptides |
AU2013258834B2 (en) | 2012-05-10 | 2017-09-07 | Zymeworks Bc Inc. | Heteromultimer constructs of immunoglobulin heavy chains with mutations in the Fc domain |
WO2013170191A1 (en) | 2012-05-11 | 2013-11-14 | Genentech, Inc. | Methods of using antagonists of nad biosynthesis from nicotinamide |
JP2015518829A (en) | 2012-05-14 | 2015-07-06 | バイオジェン・エムエイ・インコーポレイテッドBiogen MA Inc. | LINGO-2 antagonist for treatment of conditions involving motor neurons |
UY34813A (en) | 2012-05-18 | 2013-11-29 | Amgen Inc | ANTIGEN UNION PROTEINS DIRECTED AGAINST ST2 RECEIVER |
EP2666786A1 (en) | 2012-05-21 | 2013-11-27 | PAION Deutschland GmbH | Immunotherapy for intracranial hemorrhage |
MX2014014086A (en) | 2012-05-23 | 2015-01-26 | Genentech Inc | Selection method for therapeutic agents. |
US9249182B2 (en) | 2012-05-24 | 2016-02-02 | Abbvie, Inc. | Purification of antibodies using hydrophobic interaction chromatography |
BR112014029274B1 (en) | 2012-05-24 | 2022-02-15 | Mountgate Innotech (Hk) Limited | ISOLATED ANTIBODY, PHARMACEUTICAL COMPOSITION, ANTIBODY USE, AND, KIT TO TREAT RABIC INFECTION |
CA2875096A1 (en) | 2012-06-15 | 2013-12-19 | Genentech, Inc. | Anti-pcsk9 antibodies, formulations, dosing, and methods of use |
US9499634B2 (en) | 2012-06-25 | 2016-11-22 | Zymeworks Inc. | Process and methods for efficient manufacturing of highly pure asymmetric antibodies in mammalian cells |
RU2015100656A (en) | 2012-06-27 | 2016-08-20 | Ф. Хоффманн-Ля Рош Аг | METHOD FOR PRODUCING ANTIBODY FC-FRAGMENT CONNECTING, INCLUDING AT LEAST ONE CONNECTING GROUP, WHICH SPECIALLY RELATED TO THE TARGET, AND THEIR APPLICATION |
CN104395339A (en) | 2012-06-27 | 2015-03-04 | 弗·哈夫曼-拉罗切有限公司 | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
RU2644263C2 (en) | 2012-06-27 | 2018-02-08 | Ф. Хоффманн-Ля Рош Аг | Method for selection and production of selective and multispecific therapeutic molecules with specified properties, including, at least two, different target groups, and their applications |
EP3138578B1 (en) | 2012-07-04 | 2022-01-12 | F. Hoffmann-La Roche AG | Anti-theophylline antibodies and methods of use |
RU2630296C2 (en) | 2012-07-04 | 2017-09-06 | Ф. Хоффманн-Ля Рош Аг | Antibodies to biotin and application methods |
CN104428006B (en) | 2012-07-04 | 2017-09-08 | 弗·哈夫曼-拉罗切有限公司 | The antigen-antibody conjugate of covalent attachment |
CN104428416B (en) | 2012-07-05 | 2019-01-29 | 弗·哈夫曼-拉罗切有限公司 | Expression and excretory system |
BR112015000439A2 (en) | 2012-07-09 | 2017-12-19 | Genentech Inc | immunoconjugate, pharmaceutical formulation and methods of treating an individual and inhibiting proliferation |
WO2014011521A1 (en) | 2012-07-09 | 2014-01-16 | Genentech, Inc. | Immunoconjugates comprising anti - cd79b antibodies |
JP2015527318A (en) | 2012-07-09 | 2015-09-17 | ジェネンテック, インコーポレイテッド | Immune complex comprising anti-CD22 |
AU2013288930A1 (en) | 2012-07-09 | 2014-12-04 | Genentech, Inc. | Immunoconjugates comprising anti-CD79b antibodies |
AR091755A1 (en) | 2012-07-12 | 2015-02-25 | Abbvie Inc | PROTEINS OF UNION TO IL-1 |
CA2874554C (en) | 2012-07-13 | 2019-12-03 | Roche Glycart Ag | Bispecific anti-vegf/anti-ang-2 antibodies and their use in the treatment of ocular vascular diseases |
CN112587671A (en) | 2012-07-18 | 2021-04-02 | 博笛生物科技有限公司 | Targeted immunotherapy for cancer |
GB201213652D0 (en) | 2012-08-01 | 2012-09-12 | Oxford Biotherapeutics Ltd | Therapeutic and diagnostic target |
SG10201800535XA (en) | 2012-08-07 | 2018-02-27 | Roche Glycart Ag | Composition comprising two antibodies engineered to have reduced and increased effector function |
PE20150645A1 (en) | 2012-08-08 | 2015-05-11 | Roche Glycart Ag | INTERLEUQUIN 10 FUSION PROTEINS AND USES OF THEM |
US9771427B2 (en) | 2012-08-09 | 2017-09-26 | Roche Glycart Ag | ASGPR antibodies and uses thereof |
US9512214B2 (en) | 2012-09-02 | 2016-12-06 | Abbvie, Inc. | Methods to control protein heterogeneity |
CA2883272A1 (en) | 2012-09-02 | 2014-03-06 | Abbvie Inc. | Methods to control protein heterogeneity |
JP6444874B2 (en) | 2012-10-08 | 2018-12-26 | ロシュ グリクアート アーゲー | Fc-free antibody comprising two Fab fragments and methods of use |
AU2013331049B2 (en) | 2012-10-18 | 2018-11-15 | California Institute Of Technology | Broadly-neutralizing anti-HIV antibodies |
WO2014063194A1 (en) | 2012-10-23 | 2014-05-01 | The University Of Sydney | Elastic hydrogel |
KR20180008921A (en) | 2012-11-01 | 2018-01-24 | 애브비 인코포레이티드 | Anti-vegf/dll4 dual variable domain immunoglobulins and uses thereof |
EP2914621B1 (en) | 2012-11-05 | 2023-06-07 | Foundation Medicine, Inc. | Novel ntrk1 fusion molecules and uses thereof |
JP6302476B2 (en) | 2012-11-08 | 2018-03-28 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | HER3 antigen binding protein that binds to the HER3 beta hairpin |
EP3461501A1 (en) | 2012-11-13 | 2019-04-03 | F. Hoffmann-La Roche AG | Anti-hemagglutinin antibodies and methods of use |
US9914785B2 (en) | 2012-11-28 | 2018-03-13 | Zymeworks Inc. | Engineered immunoglobulin heavy chain-light chain pairs and uses thereof |
TW201425336A (en) | 2012-12-07 | 2014-07-01 | Amgen Inc | BCMA antigen binding proteins |
ES2702602T3 (en) | 2012-12-10 | 2019-03-04 | Allergan Pharmaceuticals Int Ltd | Manufacture of scalable three-dimensional elastic constructions |
US9550986B2 (en) | 2012-12-21 | 2017-01-24 | Abbvie Inc. | High-throughput antibody humanization |
JP2016510319A (en) | 2012-12-28 | 2016-04-07 | アッヴィ・インコーポレイテッド | Multivalent binding protein composition |
CA3150658A1 (en) | 2013-01-18 | 2014-07-24 | Foundation Medicine, Inc. | Methods of treating cholangiocarcinoma |
WO2014116596A1 (en) | 2013-01-22 | 2014-07-31 | Abbvie, Inc. | Methods for optimizing domain stability of binding proteins |
US20140242077A1 (en) | 2013-01-23 | 2014-08-28 | Abbvie, Inc. | Methods and compositions for modulating an immune response |
WO2014116749A1 (en) | 2013-01-23 | 2014-07-31 | Genentech, Inc. | Anti-hcv antibodies and methods of using thereof |
WO2014114595A1 (en) | 2013-01-23 | 2014-07-31 | Roche Glycart Ag | Predictive biomarker for cancer treatment with adcc-enhanced antibodies |
US9920121B2 (en) | 2013-01-25 | 2018-03-20 | Amgen Inc. | Antibodies targeting CDH19 for melanoma |
DK2951208T3 (en) | 2013-02-01 | 2020-01-13 | Kira Biotech Pty Ltd | ANTI-CD83 ANTIBODIES AND USE THEREOF |
PL2953971T3 (en) | 2013-02-07 | 2023-07-03 | Csl Limited | Il-11r binding proteins and uses thereof |
US20140228875A1 (en) | 2013-02-08 | 2014-08-14 | Nidus Medical, Llc | Surgical device with integrated visualization and cauterization |
GB201302447D0 (en) | 2013-02-12 | 2013-03-27 | Oxford Biotherapeutics Ltd | Therapeutic and diagnostic target |
MX2015010791A (en) | 2013-02-22 | 2015-11-26 | Hoffmann La Roche | Methods of treating cancer and preventing drug resistance. |
EP2961773B1 (en) | 2013-02-26 | 2019-03-20 | Roche Glycart AG | Bispecific t cell activating antigen binding molecules |
KR20150123811A (en) | 2013-02-26 | 2015-11-04 | 로슈 글리카트 아게 | Anti-mcsp antibodies |
HUE047925T2 (en) | 2013-02-26 | 2020-05-28 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules specific to cd3 and cea |
WO2014131711A1 (en) | 2013-02-26 | 2014-09-04 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
JP6416793B2 (en) | 2013-02-28 | 2018-10-31 | カプリオン プロテオミクス インコーポレーテッド | Tuberculosis biomarkers and uses thereof |
WO2014138364A2 (en) | 2013-03-06 | 2014-09-12 | Genentech, Inc. | Methods of treating and preventing cancer drug resistance |
SG11201507230PA (en) | 2013-03-12 | 2015-10-29 | Abbvie Inc | Human antibodies that bind human tnf-alpha and methods of preparing the same |
AR095399A1 (en) | 2013-03-13 | 2015-10-14 | Genentech Inc | FORMULATIONS WITH REDUCED OXIDATION, METHOD |
AR095398A1 (en) | 2013-03-13 | 2015-10-14 | Genentech Inc | FORMULATIONS WITH REDUCED OXIDATION |
RU2019137020A (en) | 2013-03-13 | 2021-01-14 | Дженентек, Инк. | REDUCED OXIDATION COMPOSITIONS |
SG10201913932VA (en) | 2013-03-13 | 2020-03-30 | Genentech Inc | Antibody formulations |
CA2901312C (en) | 2013-03-13 | 2022-09-06 | Seattle Genetics, Inc. | Activated carbon filtration for purification of benzodiazepine adcs |
ES2688895T3 (en) | 2013-03-13 | 2018-11-07 | F. Hoffmann-La Roche Ag | Formulations with reduced oxidation |
CN105228649B (en) | 2013-03-14 | 2019-01-18 | 雅培制药有限公司 | HCV Ag-Ab combination measurement is with method and used in composition therein |
US9017687B1 (en) | 2013-10-18 | 2015-04-28 | Abbvie, Inc. | Low acidic species compositions and methods for producing and using the same using displacement chromatography |
EP3299391B1 (en) | 2013-03-14 | 2019-12-04 | Genentech, Inc. | Anti-b7-h4 antibodies and immunoconjugates |
US9790478B2 (en) | 2013-03-14 | 2017-10-17 | Abbott Laboratories | HCV NS3 recombinant antigens and mutants thereof for improved antibody detection |
BR112015022576A2 (en) | 2013-03-14 | 2017-10-24 | Genentech Inc | pharmaceutical product and its use, kit and method for treating hyperproliferative dysfunction |
US8921526B2 (en) | 2013-03-14 | 2014-12-30 | Abbvie, Inc. | Mutated anti-TNFα antibodies and methods of their use |
JP2016514130A (en) | 2013-03-14 | 2016-05-19 | ノバルティス アーゲー | Antibody against Notch3 |
US9562099B2 (en) | 2013-03-14 | 2017-02-07 | Genentech, Inc. | Anti-B7-H4 antibodies and immunoconjugates |
CN113549148A (en) | 2013-03-14 | 2021-10-26 | 雅培制药有限公司 | HCV core lipid binding domain monoclonal antibodies |
JP2016516046A (en) | 2013-03-14 | 2016-06-02 | ジェネンテック, インコーポレイテッド | Methods for treating cancer and methods for preventing cancer drug resistance |
US9499614B2 (en) | 2013-03-14 | 2016-11-22 | Abbvie Inc. | Methods for modulating protein glycosylation profiles of recombinant protein therapeutics using monosaccharides and oligosaccharides |
AR095517A1 (en) | 2013-03-15 | 2015-10-21 | Genentech Inc | ANTIBODIES AGAINST THE CHEMIOATRAYENT RECEIVER EXPRESSED IN T HELPER 2 CELLS (ANTI-CRTh2) AND METHODS OF USE |
WO2014151866A1 (en) | 2013-03-15 | 2014-09-25 | Genentech, Inc. | Compositions and methods for diagnosis and treatment of hepatic cancers |
US11634502B2 (en) | 2013-03-15 | 2023-04-25 | Amgen Inc. | Heterodimeric bispecific antibodies |
BR112015023120A2 (en) | 2013-03-15 | 2017-11-21 | Genentech Inc | method for identifying an individual with a disease or dysfunction, method for predicting the responsiveness of an individual with a disease or dysfunction, method for determining the likelihood that an individual with a disease or dysfunction will exhibit benefit from treatment, method for selecting a therapy, Uses of a pd-11 Axis Binding Antagonist, Assay to Identify an Individual with a Disease, Diagnostic Kit, Method to Evaluate a Treatment Response, and Method to Monitor the Response of a Treated Individual |
US20140302037A1 (en) | 2013-03-15 | 2014-10-09 | Amgen Inc. | BISPECIFIC-Fc MOLECULES |
AU2014228172B2 (en) | 2013-03-15 | 2018-12-06 | Abbvie Deutschland Gmbh & Co.Kg | Anti-EGFR antibody drug conjugate formulations |
RU2015144020A (en) | 2013-03-15 | 2017-04-21 | Дженентек, Инк. | ENVIRONMENTS FOR CULTIVATION OF CELLS AND METHODS FOR PRODUCING ANTIBODIES |
PL2970875T3 (en) | 2013-03-15 | 2020-08-10 | F.Hoffmann-La Roche Ag | Cell culture compositions with antioxidants and methods for polypeptide production |
EA038918B1 (en) | 2013-03-15 | 2021-11-09 | Зинджения, Инк. | Peptide binding an epidermal growth factor receptor, multispecific complexes comprising peptide and antibodies and use thereof |
KR20150130451A (en) | 2013-03-15 | 2015-11-23 | 제넨테크, 인크. | Methods of treating cancer and preventing cancer drug resistance |
US10993420B2 (en) | 2013-03-15 | 2021-05-04 | Erasmus University Medical Center | Production of heavy chain only antibodies in transgenic mammals |
SG11201507432XA (en) | 2013-03-15 | 2015-10-29 | Abbvie Inc | Antibody drug conjugate (adc) purification |
CN105143258B (en) | 2013-03-15 | 2020-06-23 | Ac免疫有限公司 | anti-Tau antibodies and methods of use |
CA2904448A1 (en) | 2013-03-15 | 2014-09-18 | Tariq Ghayur | Dual specific binding proteins directed against il-1.beta. and/or il-17 |
GB201306589D0 (en) | 2013-04-11 | 2013-05-29 | Abeterno Ltd | Live cell imaging |
ES2703192T3 (en) | 2013-04-29 | 2019-03-07 | Agrosavfe Nv | Agrochemical compositions that contain antibodies that bind sphingolipids |
US11117975B2 (en) | 2013-04-29 | 2021-09-14 | Teva Pharmaceuticals Australia Pty Ltd | Anti-CD38 antibodies and fusions to attenuated interferon alpha-2B |
DK3677591T3 (en) | 2013-04-29 | 2023-03-20 | Teva Pharmaceuticals Australia Pty Ltd | Anti-CD38 antibodies and fusions to attenuated interferon alpha-2b |
KR102293064B1 (en) | 2013-05-20 | 2021-08-23 | 제넨테크, 인크. | Anti-transferrin receptor antibodies and methods of use |
AU2014273966B2 (en) | 2013-05-30 | 2017-08-31 | Kiniksa Pharmaceuticals, Ltd. | Oncostatin M receptor antigen binding proteins |
CN111518199A (en) | 2013-07-18 | 2020-08-11 | 图鲁斯生物科学有限责任公司 | Humanized antibodies with ultralong complementarity determining regions |
EP3022224A2 (en) | 2013-07-18 | 2016-05-25 | Fabrus, Inc. | Antibodies with ultralong complementarity determining regions |
KR102062784B1 (en) | 2013-07-23 | 2020-01-07 | 바이오콘 리미티드 | Methods for controlling fucosylation levels in proteins |
TWI623551B (en) | 2013-08-02 | 2018-05-11 | 輝瑞大藥廠 | Anti-cxcr4 antibodies and antibody-drug conjugates |
JP6382311B2 (en) | 2013-08-13 | 2018-08-29 | エラスタジェン・プロプライエタリー・リミテッドElastagen Pty Ltd | Regeneration of damaged tissue |
RU2016109247A (en) | 2013-09-17 | 2017-10-19 | Дженентек, Инк. | WAYS OF APPLICATION OF ANTIBODIES TO LGR5 |
KR102331663B1 (en) | 2013-09-27 | 2021-11-25 | 제넨테크, 인크. | Anti-pdl1 antibody formulations |
WO2015050959A1 (en) | 2013-10-01 | 2015-04-09 | Yale University | Anti-kit antibodies and methods of use thereof |
AU2014329609B2 (en) | 2013-10-02 | 2019-09-12 | Humabs Biomed Sa | Neutralizing anti-influenza A antibodies and uses thereof |
WO2015051293A2 (en) | 2013-10-04 | 2015-04-09 | Abbvie, Inc. | Use of metal ions for modulation of protein glycosylation profiles of recombinant proteins |
EP3055328A1 (en) | 2013-10-11 | 2016-08-17 | F. Hoffmann-La Roche AG | Nsp4 inhibitors and methods of use |
JP6422956B2 (en) | 2013-10-11 | 2018-11-14 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Multispecific domain exchange common variable light chain antibody |
US9181337B2 (en) | 2013-10-18 | 2015-11-10 | Abbvie, Inc. | Modulated lysine variant species compositions and methods for producing and using the same |
US8946395B1 (en) | 2013-10-18 | 2015-02-03 | Abbvie Inc. | Purification of proteins using hydrophobic interaction chromatography |
CN105744954B (en) | 2013-10-18 | 2021-03-05 | 豪夫迈·罗氏有限公司 | anti-RSPO 2 and/or anti-RSPO 3 antibodies and uses thereof |
US9085618B2 (en) | 2013-10-18 | 2015-07-21 | Abbvie, Inc. | Low acidic species compositions and methods for producing and using the same |
SG11201603127WA (en) | 2013-10-23 | 2016-05-30 | Genentech Inc | Methods of diagnosing and treating eosinophilic disorders |
US10203327B2 (en) | 2013-11-05 | 2019-02-12 | Novartis Ag | Organic compounds |
WO2015073884A2 (en) | 2013-11-15 | 2015-05-21 | Abbvie, Inc. | Glycoengineered binding protein compositions |
MY176237A (en) | 2013-11-21 | 2020-07-24 | Hoffmann La Roche | Anti-alpha-synuclein antibodies and methods of use |
CA2931356A1 (en) | 2013-11-27 | 2015-06-04 | Zymeworks Inc. | Bispecific antigen-binding constructs targeting her2 |
JP6612246B2 (en) | 2013-11-28 | 2019-11-27 | シーエスエル、リミテッド | How to treat nephropathy |
TW201536320A (en) | 2013-12-02 | 2015-10-01 | Abbvie Inc | Compositions and methods for treating osteoarthritis |
US9309314B2 (en) | 2013-12-03 | 2016-04-12 | Agency For Science, Technology And Research (A*Star) | Polypeptides, nucleic acids and uses thereof |
CN105916519B (en) | 2013-12-09 | 2021-10-22 | 爱乐科斯公司 | anti-Siglec-8 antibodies and methods of use thereof |
EP3080164B1 (en) | 2013-12-13 | 2019-01-16 | Genentech, Inc. | Anti-cd33 antibodies and immunoconjugates |
EP2883883A1 (en) | 2013-12-16 | 2015-06-17 | Cardio3 Biosciences S.A. | Therapeutic targets and agents useful in treating ischemia reperfusion injury |
MX2016007965A (en) | 2013-12-17 | 2016-10-28 | Genentech Inc | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists. |
SG10201809385RA (en) | 2013-12-17 | 2018-11-29 | Genentech Inc | Anti-cd3 antibodies and methods of use |
SG11201604875PA (en) | 2013-12-17 | 2016-07-28 | Genentech Inc | Methods of treating cancer using pd-1 axis binding antagonists and an anti-cd20 antibody |
KR102630750B1 (en) | 2013-12-17 | 2024-01-30 | 제넨테크, 인크. | Methods of treating cancers using pd-1 axis binding antagonists and taxanes |
ES2851386T3 (en) | 2013-12-18 | 2021-09-06 | Csl Ltd | Wound treatment method |
CN105899533B (en) | 2013-12-20 | 2019-10-11 | 豪夫迈·罗氏有限公司 | Anti- Tau (pS422) antibody and application method of humanization |
TWI670283B (en) | 2013-12-23 | 2019-09-01 | 美商建南德克公司 | Antibodies and methods of use |
MX2016008189A (en) | 2014-01-03 | 2016-09-29 | Hoffmann La Roche | Covalently linked helicar-anti-helicar antibody conjugates and uses thereof. |
MX2016008187A (en) | 2014-01-03 | 2016-09-29 | Hoffmann La Roche | Bispecific anti-hapten/anti-blood brain barrier receptor antibodies, complexes thereof and their use as blood brain barrier shuttles. |
JP6521464B2 (en) | 2014-01-03 | 2019-05-29 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Covalently linked polypeptide toxin-antibody conjugates |
CA2932547C (en) | 2014-01-06 | 2023-05-23 | F. Hoffmann-La Roche Ag | Monovalent blood brain barrier shuttle modules |
EP3092254A4 (en) | 2014-01-10 | 2017-09-20 | Birdie Biopharmaceuticals Inc. | Compounds and compositions for treating her2 positive tumors |
EP3096797A1 (en) | 2014-01-24 | 2016-11-30 | F. Hoffmann-La Roche AG | Methods of using anti-steap1 antibodies and immunoconjugates |
EP3102197B1 (en) | 2014-02-04 | 2018-08-29 | Genentech, Inc. | Mutant smoothened and methods of using the same |
MX2016010237A (en) | 2014-02-08 | 2017-04-27 | Genentech Inc | Methods of treating alzheimer's disease. |
EP3718563A1 (en) | 2014-02-08 | 2020-10-07 | F. Hoffmann-La Roche AG | Methods of treating alzheimer's disease |
CA2936565C (en) | 2014-02-12 | 2020-08-11 | Genentech, Inc. | Anti-jagged1 antibodies and methods of use |
JP6619745B2 (en) | 2014-02-20 | 2019-12-11 | アラーガン、インコーポレイテッドAllergan,Incorporated | Complement component C5 antibody |
CN106029693A (en) | 2014-02-21 | 2016-10-12 | 豪夫迈·罗氏有限公司 | Anti-IL-13/IL-17 bispecific antibodies and uses thereof |
US9796776B2 (en) | 2014-02-27 | 2017-10-24 | Allergan, Inc. | Complement factor Bb antibodies |
EP3110446B1 (en) | 2014-02-28 | 2021-12-01 | Allakos Inc. | Methods and compositions for treating siglec-8 associated diseases |
ES2897765T3 (en) | 2014-03-14 | 2022-03-02 | Hoffmann La Roche | Methods and compositions for the secretion of heterologous polypeptides |
KR20220123560A (en) | 2014-03-21 | 2022-09-07 | 애브비 인코포레이티드 | Anti-egfr antibodies and antibody drug conjugates |
WO2015140591A1 (en) | 2014-03-21 | 2015-09-24 | Nordlandssykehuset Hf | Anti-cd14 antibodies and uses thereof |
KR20160137599A (en) | 2014-03-24 | 2016-11-30 | 제넨테크, 인크. | Cancer treatment with c-met antagonists and correlation of the latter with hgf expression |
AU2015241038A1 (en) | 2014-03-31 | 2016-10-13 | Genentech, Inc. | Combination therapy comprising anti-angiogenesis agents and OX40 binding agonists |
PT3126394T (en) | 2014-03-31 | 2019-12-19 | Hoffmann La Roche | Anti-ox40 antibodies and methods of use |
BR112016024319B1 (en) | 2014-04-18 | 2024-01-23 | Acceleron Pharma Inc | USE OF A COMPOSITION COMPRISING AN ActRII ANTAGONIST FOR THE MANUFACTURING OF A MEDICATION FOR TREATING OR PREVENTING A COMPLICATION OF SICKLE CELL ANEMIA |
WO2015164615A1 (en) | 2014-04-24 | 2015-10-29 | University Of Oslo | Anti-gluten antibodies and uses thereof |
UA119352C2 (en) | 2014-05-01 | 2019-06-10 | Тева Фармасьютикалз Острейліа Пті Лтд | Combination of lenalidomide or pomalidomide and cd38 antibody-attenuated interferon-alpha constructs, and the use thereof |
US11474101B2 (en) | 2014-05-08 | 2022-10-18 | Novodiax, Inc. | Direct immunohistochemistry assay |
EP3888690A3 (en) | 2014-05-16 | 2021-10-20 | MedImmune, LLC | Molecules with altered neonate fc receptor binding having enhanced therapeutic and diagnostic properties |
JP2017522861A (en) | 2014-05-22 | 2017-08-17 | ジェネンテック, インコーポレイテッド | Anti-GPC3 antibody and immunoconjugate |
EP3146071B1 (en) | 2014-05-23 | 2020-09-02 | F. Hoffmann-La Roche AG | Mit biomarkers and methods using the same |
WO2015191617A2 (en) | 2014-06-09 | 2015-12-17 | Biomed Valley Discoveries, Inc. | Combination therapies using anti-metabolites and agents that target tumor-associated stroma or tumor cells |
US10434174B2 (en) | 2014-06-09 | 2019-10-08 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Combination therapies using platinum agents and agents that target tumor-associated stroma or tumor cells |
WO2015191590A2 (en) | 2014-06-09 | 2015-12-17 | Biomed Valley Discoveries, Inc. | Combination therapies targeting tumor-associated stroma or tumor cells and microtubules |
WO2015191610A2 (en) | 2014-06-09 | 2015-12-17 | Biomed Valley Discoveries, Inc. | Combination therapies using agents that target tumor-associated stroma or tumor cells and other pathways |
US10799584B2 (en) | 2014-06-09 | 2020-10-13 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services National Institutes Of Health | Combination therapies using agents that target tumor-associated stroma or tumor cells and alkylating agents |
US10758614B2 (en) | 2014-06-09 | 2020-09-01 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services National Institutes Of Health | Combination therapies targeting tumor-associated stroma or tumor cells and topoisomerase |
US11034757B2 (en) | 2014-06-09 | 2021-06-15 | Biomed Valley Discoveries, Inc. | Combination therapies using agents that target tumor-associated stroma or tumor cells and tumor vasculature |
CA2949982A1 (en) | 2014-06-11 | 2015-12-17 | Genentech, Inc. | Anti-lgr5 antibodies and uses thereof |
BR122023023170A2 (en) | 2014-06-13 | 2024-02-20 | Acceleron Pharma Inc. | USE OF AN ACTRII ANTAGONIST IN THE TREATMENT OR PREVENTION OF SKIN ULCERS ASSOCIATED WITH BETA-THALASSEMIA |
WO2015191986A1 (en) | 2014-06-13 | 2015-12-17 | Genentech, Inc. | Methods of treating and preventing cancer drug resistance |
NL2013007B1 (en) | 2014-06-16 | 2016-07-05 | Ablynx Nv | Methods of treating TTP with immunoglobulin single variable domains and uses thereof. |
NL2013661B1 (en) | 2014-10-21 | 2016-10-05 | Ablynx Nv | KV1.3 Binding immunoglobulins. |
MX2016015280A (en) | 2014-06-26 | 2017-03-03 | Hoffmann La Roche | Anti-brdu antibodies and methods of use. |
AR100978A1 (en) | 2014-06-26 | 2016-11-16 | Hoffmann La Roche | ANTI-Tau HUMANIZED ANTIBODY BRAIN LAUNCHERS (pS422) AND USES OF THE SAME |
SI3164492T1 (en) | 2014-07-03 | 2020-02-28 | F. Hoffmann-La Roche Ag | Polypeptide expression systems |
CN112546230A (en) | 2014-07-09 | 2021-03-26 | 博笛生物科技有限公司 | Combination therapeutic compositions and combination therapeutic methods for treating cancer |
CA2954446A1 (en) | 2014-07-09 | 2016-01-14 | Shanghai Birdie Biotech, Inc. | Anti-pd-l1 combinations for treating tumors |
MX2017000363A (en) | 2014-07-11 | 2017-04-27 | Genentech Inc | Notch pathway inhibition. |
KR102360693B1 (en) | 2014-07-11 | 2022-02-08 | 벤타나 메디컬 시스템즈, 인코포레이티드 | Anti-pd-l1 antibodies and diagnostic uses thereof |
WO2016009436A1 (en) | 2014-07-15 | 2016-01-21 | Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. | Isolated polypeptides of cd44 and uses thereof |
MX2017001556A (en) | 2014-08-04 | 2017-05-15 | Hoffmann La Roche | Bispecific t cell activating antigen binding molecules. |
BR112017003835A2 (en) | 2014-08-28 | 2018-04-10 | Bioatla Llc | chimeric antigen receptor, expression vector, genetically engineered cytotoxic cell, pharmaceutical composition, and methods for producing a chimeric antigen receptor and for treating a disease in an individual. |
CN112587672A (en) | 2014-09-01 | 2021-04-02 | 博笛生物科技有限公司 | anti-PD-L1 conjugates for the treatment of tumors |
US9751946B2 (en) | 2014-09-12 | 2017-09-05 | Genentech, Inc. | Anti-CLL-1 antibodies and immunoconjugates |
CN113698488A (en) | 2014-09-12 | 2021-11-26 | 基因泰克公司 | anti-B7-H4 antibodies and immunoconjugates |
BR112017004631A2 (en) | 2014-09-12 | 2018-01-30 | Genentech, Inc. | antibody, nucleic acid, host cell, antibody production method, immunoconjugate, pharmaceutical formulation, methods of treatment, cell proliferation inhibition and human her2 detection and method for cancer detection |
MX2017003121A (en) | 2014-09-15 | 2017-08-02 | Genentech Inc | Antibody formulations. |
WO2016044396A1 (en) | 2014-09-17 | 2016-03-24 | Genentech, Inc. | Immunoconjugates comprising anti-her2 antibodies and pyrrolobenzodiazepines |
SI3262071T1 (en) | 2014-09-23 | 2020-07-31 | F. Hoffmann-La Roche Ag | Method of using anti-cd79b immunoconjugates |
DK3204095T3 (en) | 2014-10-10 | 2019-07-15 | Ablynx Nv | INHALATION APPLICATION FOR USE IN AEROSOL TREATMENT OF AIR ROAD DISEASES |
US10561805B2 (en) | 2014-10-10 | 2020-02-18 | Ablynx N.V. | Methods of treating RSV infections |
US9732148B2 (en) | 2014-10-16 | 2017-08-15 | Genentech, Inc. | Anti-α-synuclein antibodies and methods of use |
WO2016059602A2 (en) | 2014-10-16 | 2016-04-21 | Glaxo Group Limited | Methods of treating cancer and related compositions |
AU2015336946A1 (en) | 2014-10-23 | 2017-04-13 | La Trobe University | Fn14-binding proteins and uses thereof |
EP3209695A4 (en) | 2014-10-23 | 2018-05-30 | DendroCyte BioTech Pty Ltd | Cd83 binding proteins and uses thereof |
HUE054075T2 (en) | 2014-10-29 | 2021-08-30 | Seagen Inc | Dosage and administration of non-fucosylated anti-cd40 antibodies |
SG11201703251TA (en) | 2014-10-29 | 2017-05-30 | Teva Pharmaceuticals Australia Pty Ltd | INTERFERON α2B VARIANTS |
AU2015343339A1 (en) | 2014-11-03 | 2017-06-15 | Genentech, Inc. | Methods and biomarkers for predicting efficacy and evaluation of an OX40 agonist treatment |
CN106796235B (en) | 2014-11-03 | 2021-01-29 | 豪夫迈·罗氏有限公司 | Assays for detecting T cell immune subsets and methods of use thereof |
CA2966548A1 (en) | 2014-11-05 | 2016-05-12 | Agrosavfe Nv | Transgenic plant comprising a polynucleotide encoding a variable domain of heavy-chain antibody |
KR102544705B1 (en) | 2014-11-05 | 2023-06-15 | 제넨테크, 인크. | Methods of producing two chain proteins in bacteria |
JP6875276B2 (en) | 2014-11-05 | 2021-05-19 | ジェネンテック, インコーポレイテッド | Method of producing double-stranded protein in bacteria |
RU2017119428A (en) | 2014-11-06 | 2018-12-06 | Дженентек, Инк. | COMBINED THERAPY, INCLUDING THE USE OF OX40-CONNECTING AGONISTS AND TIGIT INHIBITORS |
WO2016073157A1 (en) | 2014-11-06 | 2016-05-12 | Genentech, Inc. | Anti-ang2 antibodies and methods of use thereof |
CN107105632A (en) | 2014-11-10 | 2017-08-29 | 豪夫迈·罗氏有限公司 | Nephrosis animal model and its therapeutic agent |
EA201791029A1 (en) | 2014-11-10 | 2017-12-29 | Дженентек, Инк. | ANTIBODIES AGAINST INTERLEUKIN-33 AND THEIR APPLICATION |
TWI713474B (en) | 2014-11-14 | 2020-12-21 | 瑞士商赫孚孟拉羅股份公司 | Antigen binding molecules comprising a tnf family ligand trimer |
CN107429075B (en) | 2014-11-17 | 2022-11-01 | 卡内基梅隆大学 | Activatable two-component photosensitizer |
JP2017537090A (en) | 2014-11-17 | 2017-12-14 | ジェネンテック, インコーポレイテッド | Combination therapy comprising OX40 binding agonist and PD-1 axis binding antagonist |
WO2016081640A1 (en) | 2014-11-19 | 2016-05-26 | Genentech, Inc. | Anti-transferrin receptor / anti-bace1 multispecific antibodies and methods of use |
CN107108745B (en) | 2014-11-19 | 2021-01-12 | 基因泰克公司 | Antibodies against BACE1 and their use for immunotherapy of neurological diseases |
US10508151B2 (en) | 2014-11-19 | 2019-12-17 | Genentech, Inc. | Anti-transferrin receptor antibodies and methods of use |
EP3747905A1 (en) | 2014-11-20 | 2020-12-09 | F. Hoffmann-La Roche AG | Common light chains and methods of use |
KR20240024318A (en) | 2014-11-20 | 2024-02-23 | 에프. 호프만-라 로슈 아게 | Combination therapy of t cell activating bispecific antigen binding molecules cd3 abd folate receptor 1 (folr1) and pd-1 axis binding antagonists |
CN107001482B (en) | 2014-12-03 | 2021-06-15 | 豪夫迈·罗氏有限公司 | Multispecific antibodies |
MA41119A (en) | 2014-12-03 | 2017-10-10 | Acceleron Pharma Inc | METHODS OF TREATMENT OF MYELODYSPLASIC SYNDROMES AND SIDEROBLASTIC ANEMIA |
CN107108739B (en) | 2014-12-05 | 2022-01-04 | 豪夫迈·罗氏有限公司 | anti-CD 79b antibodies and methods of use |
BR112017011234A2 (en) | 2014-12-10 | 2018-03-27 | Genentech Inc | antibodies to the blood-brain barrier receptor and methods of use |
US10093733B2 (en) | 2014-12-11 | 2018-10-09 | Abbvie Inc. | LRP-8 binding dual variable domain immunoglobulin proteins |
CA2971278C (en) | 2014-12-19 | 2023-09-19 | Ablynx N.V. | Cysteine linked nanobody dimers |
SG10201710322VA (en) | 2014-12-19 | 2018-02-27 | Chugai Pharmaceutical Co Ltd | Anti-c5 antibodies and methods of use |
WO2016094962A1 (en) | 2014-12-19 | 2016-06-23 | Monash University | Il-21 antibodies |
WO2016112270A1 (en) | 2015-01-08 | 2016-07-14 | Biogen Ma Inc. | Lingo-1 antagonists and uses for treatment of demyelinating disorders |
AU2016206191B2 (en) | 2015-01-09 | 2017-08-03 | Adalta Limited | CXCR4 binding molecules |
WO2016117346A1 (en) | 2015-01-22 | 2016-07-28 | Chugai Seiyaku Kabushiki Kaisha | A combination of two or more anti-c5 antibodies and methods of use |
US11304676B2 (en) | 2015-01-23 | 2022-04-19 | The University Of North Carolina At Chapel Hill | Apparatuses, systems, and methods for preclinical ultrasound imaging of subjects |
JP6871866B2 (en) | 2015-02-03 | 2021-05-19 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Anti-RHO GTPASE conformational single domain antibody and its use |
US10330683B2 (en) | 2015-02-04 | 2019-06-25 | Genentech, Inc. | Mutant smoothened and methods of using the same |
CN114773469A (en) | 2015-02-05 | 2022-07-22 | 中外制药株式会社 | Antibodies comprising an ion concentration-dependent antigen-binding domain, FC region variants, IL-8-binding antibodies and uses thereof |
US10457737B2 (en) | 2015-02-09 | 2019-10-29 | Research Development Foundation | Engineered immunoglobulin Fc polypeptides displaying improved complement activation |
US20170151281A1 (en) | 2015-02-19 | 2017-06-01 | Batu Biologics, Inc. | Chimeric antigen receptor dendritic cell (car-dc) for treatment of cancer |
WO2016135041A1 (en) | 2015-02-26 | 2016-09-01 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Fusion proteins and antibodies comprising thereof for promoting apoptosis |
CN107430117A (en) | 2015-03-16 | 2017-12-01 | 豪夫迈·罗氏有限公司 | Detection and quantitative IL 13 method and the purposes in diagnosing and treating Th2 relevant diseases |
WO2016146833A1 (en) | 2015-03-19 | 2016-09-22 | F. Hoffmann-La Roche Ag | Biomarkers for nad(+)-diphthamide adp ribosyltransferase resistance |
MA54328A (en) | 2015-04-06 | 2021-10-06 | Acceleron Pharma Inc | TYPE I AND TYPE II RECEPTOR HETEROMULTIMERS OF THE TGF-BETA SUPERFAMILY AND THEIR USES |
MA41919A (en) | 2015-04-06 | 2018-02-13 | Acceleron Pharma Inc | ALK4 HETEROMULTIMERS: ACTRIIB AND THEIR USES |
DK3280441T3 (en) | 2015-04-07 | 2021-11-15 | Alector Llc | ANTI-SORTILINE ANTIBODIES AND PROCEDURES FOR USE |
CA2981183A1 (en) | 2015-04-07 | 2016-10-13 | Greg Lazar | Antigen binding complex having agonistic activity and methods of use |
WO2016172160A1 (en) | 2015-04-21 | 2016-10-27 | Genentech, Inc. | Compositions and methods for prostate cancer analysis |
JP7044553B2 (en) | 2015-04-24 | 2022-03-30 | ジェネンテック, インコーポレイテッド | How to identify bacteria containing bound polypeptides |
BR112017023158A2 (en) | 2015-04-30 | 2018-07-24 | Harvard College | anti-ap2 antibodies and antigen binding agents to treat metabolic disorders |
EP3778640A1 (en) | 2015-05-01 | 2021-02-17 | Genentech, Inc. | Masked anti-cd3 antibodies and methods of use |
US20160346387A1 (en) | 2015-05-11 | 2016-12-01 | Genentech, Inc. | Compositions and methods of treating lupus nephritis |
DK3294770T3 (en) | 2015-05-12 | 2020-12-07 | Hoffmann La Roche | Therapeutic and diagnostic methods for cancer |
EP3095465A1 (en) | 2015-05-19 | 2016-11-23 | U3 Pharma GmbH | Combination of fgfr4-inhibitor and bile acid sequestrant |
CN107771182A (en) | 2015-05-29 | 2018-03-06 | 豪夫迈·罗氏有限公司 | The anti-Ebola virus glycoproteins antibody of humanization and application method |
IL294138A (en) | 2015-05-29 | 2022-08-01 | Genentech Inc | Therapeutic and diagnostic methods for cancer |
RS59935B1 (en) | 2015-05-29 | 2020-03-31 | Abbvie Inc | Anti-cd40 antibodies and uses thereof |
KR20180011117A (en) | 2015-05-31 | 2018-01-31 | 큐어제닉스 코포레이션 | Composite composition for immunotherapy |
EP3302552A1 (en) | 2015-06-02 | 2018-04-11 | H. Hoffnabb-La Roche Ag | Compositions and methods for using anti-il-34 antibodies to treat neurological diseases |
TWI790642B (en) | 2015-06-05 | 2023-01-21 | 美商建南德克公司 | Anti-tau antibodies and methods of use |
JP2018521019A (en) | 2015-06-08 | 2018-08-02 | ジェネンテック, インコーポレイテッド | Method of treating cancer using anti-OX40 antibody |
AU2016274584A1 (en) | 2015-06-08 | 2018-01-04 | Genentech, Inc. | Methods of treating cancer using anti-OX40 antibodies and PD-1 axis binding antagonists |
JP2018518491A (en) | 2015-06-12 | 2018-07-12 | アレクトル エルエルシー | Anti-CD33 antibody and method of use thereof |
SG10201912085WA (en) | 2015-06-12 | 2020-02-27 | Alector Llc | Anti-cd33 antibodies and methods of use thereof |
AR104987A1 (en) | 2015-06-15 | 2017-08-30 | Genentech Inc | ANTIBODY-DRUG IMMUNOCUJADOS UNITED BY NON-PEPTIDIC LINKERS |
TW201710286A (en) | 2015-06-15 | 2017-03-16 | 艾伯維有限公司 | Binding proteins against VEGF, PDGF, and/or their receptors |
CN114507289A (en) | 2015-06-16 | 2022-05-17 | 豪夫迈·罗氏有限公司 | Humanized and affinity matured antibodies to FcRH5 and methods of use |
JP6996983B2 (en) | 2015-06-16 | 2022-02-21 | ジェネンテック, インコーポレイテッド | Anti-CLL-1 antibody and how to use |
WO2016204966A1 (en) | 2015-06-16 | 2016-12-22 | Genentech, Inc. | Anti-cd3 antibodies and methods of use |
US10774145B2 (en) | 2015-06-17 | 2020-09-15 | Allakos Inc. | Methods and compositions for treating fibrotic diseases |
CN107787331B (en) | 2015-06-17 | 2022-01-11 | 豪夫迈·罗氏有限公司 | anti-HER 2 antibodies and methods of use |
KR20180018538A (en) | 2015-06-17 | 2018-02-21 | 제넨테크, 인크. | Methods for the treatment of locally advanced or metastatic breast cancer using PD-1 axis-binding antagonists and taxanes |
EP3744732A1 (en) | 2015-06-24 | 2020-12-02 | F. Hoffmann-La Roche AG | Humanized anti-tau(ps422) antibodies and methods of use |
US9861621B2 (en) | 2015-06-29 | 2018-01-09 | Biomed Valley Discoveries, Inc. | LPT-723 and immune checkpoint inhibitor combinations and methods of treatment |
CA2989936A1 (en) | 2015-06-29 | 2017-01-05 | Genentech, Inc. | Type ii anti-cd20 antibody for use in organ transplantation |
CA3162816A1 (en) | 2015-06-29 | 2017-01-05 | Ventana Medical Systems, Inc. | Materials and methods for performing histochemical assays for human pro-epiregulin and amphiregulin |
WO2017004330A1 (en) | 2015-06-30 | 2017-01-05 | Seattle Genetics, Inc. | Anti-ntb-a antibodies and related compositions and methods |
WO2017023866A1 (en) | 2015-07-31 | 2017-02-09 | Boston Biomedical, Inc. | Method of targeting stat3 and other non-druggable proteins |
TWI797060B (en) | 2015-08-04 | 2023-04-01 | 美商再生元醫藥公司 | Taurine supplemented cell culture medium and methods of use |
CN108348578B (en) | 2015-08-04 | 2022-08-09 | 阿塞勒隆制药公司 | Methods for treating myeloproliferative disorders |
SG10201913625XA (en) | 2015-08-07 | 2020-03-30 | Imaginab Inc | Antigen binding constructs to target molecules |
CN105384825B (en) | 2015-08-11 | 2018-06-01 | 南京传奇生物科技有限公司 | A kind of bispecific chimeric antigen receptor and its application based on single domain antibody |
EP3932953A1 (en) | 2015-08-28 | 2022-01-05 | F. Hoffmann-La Roche AG | Anti-hypusine antibodies and uses thereof |
WO2017046335A1 (en) | 2015-09-18 | 2017-03-23 | INSERM (Institut National de la Santé et de la Recherche Médicale) | T cell receptors (tcr) and uses thereof for the diagnosis and treatment of diabetes |
WO2017046994A1 (en) | 2015-09-18 | 2017-03-23 | Chugai Seiyaku Kabushiki Kaisha | Il-8-binding antibodies and uses thereof |
EP3353206A1 (en) | 2015-09-22 | 2018-08-01 | Spring Bioscience Corporation | Anti-ox40 antibodies and diagnostic uses thereof |
CN116987187A (en) | 2015-09-23 | 2023-11-03 | 豪夫迈·罗氏有限公司 | Optimized variants of anti-VEGF antibodies |
CN108289954B (en) | 2015-09-24 | 2022-05-31 | 阿布维特罗有限责任公司 | HIV antibody compositions and methods of use |
WO2017055484A1 (en) | 2015-09-29 | 2017-04-06 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for determining the metabolic status of lymphomas |
AR106188A1 (en) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE |
MA43345A (en) | 2015-10-02 | 2018-08-08 | Hoffmann La Roche | PYRROLOBENZODIAZEPINE ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
BR112018005164A2 (en) | 2015-10-02 | 2019-10-01 | Hoffmann La Roche | antibodies, nucleic acid, host cell, method for producing an antibody, pharmaceutical formulation, use of the antibody and method of treating an individual who has cancer |
JP6937746B2 (en) | 2015-10-02 | 2021-09-22 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Bispecific anti-CD19 × CD3T cell-activating antigen-binding molecule |
WO2017055392A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xcd44v6 bispecific t cell activating antigen binding molecules |
EP3356409A2 (en) | 2015-10-02 | 2018-08-08 | H. Hoffnabb-La Roche Ag | Bispecific t cell activating antigen binding molecules |
WO2017055385A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xgd2 bispecific t cell activating antigen binding molecules |
KR102431342B1 (en) | 2015-10-02 | 2022-08-10 | 에프. 호프만-라 로슈 아게 | Bispecific antibodies specific for pd1 and tim3 |
WO2017055395A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xrob04 bispecific t cell activating antigen binding molecules |
JP2018536389A (en) | 2015-10-02 | 2018-12-13 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Bispecific cell-activating antigen binding molecule that binds mesothelin and CD3 |
WO2017055393A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xtim-3 bispecific t cell activating antigen binding molecules |
MA43017A (en) | 2015-10-02 | 2018-08-08 | Hoffmann La Roche | BISPECIFIC ANTIBODIES SPECIFIC TO A TNF CO-STIMULATION RECEPTOR |
EP3356410B1 (en) | 2015-10-02 | 2021-10-20 | F. Hoffmann-La Roche AG | Bispecific anti-ceaxcd3 t cell activating antigen binding molecules |
CN108271377B (en) | 2015-10-07 | 2021-11-19 | 豪夫迈·罗氏有限公司 | Bispecific antibodies having a tetravalent targeting co-stimulatory TNF receptor |
US10556953B2 (en) | 2015-10-12 | 2020-02-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Agent capable of depleting CD8 T cells for the treatment of myocardial infarction or acute myocardial infarction |
MA43354A (en) | 2015-10-16 | 2018-08-22 | Genentech Inc | CONJUGATE DRUG CONJUGATES WITH CLOUDY DISULPHIDE |
MA45326A (en) | 2015-10-20 | 2018-08-29 | Genentech Inc | CALICHEAMICIN-ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
US10604577B2 (en) | 2015-10-22 | 2020-03-31 | Allakos Inc. | Methods and compositions for treating systemic mastocytosis |
JP6949016B2 (en) | 2015-10-29 | 2021-10-13 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Antivariant Fc region antibody and usage |
EP3184547A1 (en) | 2015-10-29 | 2017-06-28 | F. Hoffmann-La Roche AG | Anti-tpbg antibodies and methods of use |
SI3368578T1 (en) | 2015-10-30 | 2021-08-31 | F. Hoffmann-La Roche Ag | Anti-htra1 antibodies and methods of use thereof |
WO2017075173A2 (en) | 2015-10-30 | 2017-05-04 | Genentech, Inc. | Anti-factor d antibodies and conjugates |
CN108602884A (en) | 2015-11-08 | 2018-09-28 | 豪夫迈·罗氏有限公司 | The method for screening multi-specificity antibody |
EP3374389A1 (en) | 2015-11-13 | 2018-09-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti- nkg2d single domain antibodies and uses thereof |
KR20180083402A (en) | 2015-11-20 | 2018-07-20 | 오스트레일리언 바이오메디컬 컴퍼니 피티와이 리미티드 | Compounds for medical use |
WO2017091706A1 (en) | 2015-11-23 | 2017-06-01 | Acceleron Pharma Inc. | Methods for treating eye disorders |
EP3932945A1 (en) | 2015-11-27 | 2022-01-05 | Ablynx NV | Polypeptides inhibiting cd40l |
EP3178848A1 (en) | 2015-12-09 | 2017-06-14 | F. Hoffmann-La Roche AG | Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies |
MX2018005229A (en) | 2015-12-09 | 2019-04-29 | F HoffmannLa Roche Ag | Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies. |
TWI597292B (en) | 2015-12-18 | 2017-09-01 | 中外製藥股份有限公司 | Anti-c5 antibodies and methods of use |
JP6970090B2 (en) | 2015-12-18 | 2021-11-24 | エーザイ・アール・アンド・ディー・マネジメント株式会社 | Immunoglobulin conjugated with C-terminal lysine |
MX2018008063A (en) | 2015-12-30 | 2018-11-29 | Genentech Inc | Use of tryptophan derivatives for protein formulations. |
CN115400220A (en) | 2015-12-30 | 2022-11-29 | 豪夫迈·罗氏有限公司 | Preparation for reducing degradation of polysorbate |
US20200264165A1 (en) | 2016-01-04 | 2020-08-20 | Inserm (Institut National De La Sante Et De Larecherche Medicale) | Use of pd-1 and tim-3 as a measure for cd8+ cells in predicting and treating renal cell carcinoma |
WO2017118307A1 (en) | 2016-01-05 | 2017-07-13 | 江苏恒瑞医药股份有限公司 | Pcsk9 antibody, antigen-binding fragment thereof, and medical uses thereof |
KR20180097615A (en) | 2016-01-08 | 2018-08-31 | 에프. 호프만-라 로슈 아게 | Methods for the treatment of CEA-positive cancers using PD-1 axis-binding antagonists and anti-CEA / anti-CD3 bispecific antibodies |
CA3011739A1 (en) | 2016-01-20 | 2017-07-27 | Genentech, Inc. | High dose treatments for alzheimer's disease |
WO2017129558A1 (en) | 2016-01-25 | 2017-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for predicting or treating myelopoiesis-driven cardiometabolic diseases and sepsis |
ES2924775T3 (en) | 2016-01-28 | 2022-10-10 | Inst Nat Sante Rech Med | Methods and pharmaceutical composition for the treatment of cancer |
ES2924741T3 (en) | 2016-01-28 | 2022-10-10 | Inst Nat Sante Rech Med | Methods to Increase the Potency of Immune Checkpoint Inhibitors |
WO2017129763A1 (en) | 2016-01-28 | 2017-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of signet ring cell gastric cancer |
AU2017213826A1 (en) | 2016-02-04 | 2018-08-23 | Curis, Inc. | Mutant smoothened and methods of using the same |
MX2018010361A (en) | 2016-02-29 | 2019-07-08 | Genentech Inc | Therapeutic and diagnostic methods for cancer. |
MY189113A (en) | 2016-03-02 | 2022-01-26 | Eisai R&D Man Co Ltd | Eribulin-based antibody-drug conjugates and methods of use |
US11027021B2 (en) | 2016-03-15 | 2021-06-08 | Seagen Inc. | Combinations of PBD-based antibody drug conjugates with Bcl-2 inhibitors |
EP3430054B1 (en) | 2016-03-15 | 2021-12-29 | Chugai Seiyaku Kabushiki Kaisha | Methods of treating cancers using pd-1 axis binding antagonists and anti-gpc3 antibodies |
PT3433280T (en) | 2016-03-22 | 2023-06-15 | Hoffmann La Roche | Protease-activated t cell bispecific molecules |
SI3433280T1 (en) | 2016-03-22 | 2023-07-31 | F. Hoffmann-La Roche Ag | Protease-activated t cell bispecific molecules |
US20170315132A1 (en) | 2016-03-25 | 2017-11-02 | Genentech, Inc. | Multiplexed total antibody and antibody-conjugated drug quantification assay |
KR20240034883A (en) | 2016-03-29 | 2024-03-14 | 얀센 바이오테크 인코포레이티드 | Treating psoriasis with increased interval dosing of anti-il12 and/or -23 antibody |
EP3231813A1 (en) | 2016-03-29 | 2017-10-18 | F. Hoffmann-La Roche AG | Trimeric costimulatory tnf family ligand-containing antigen binding molecules |
US20200330459A1 (en) | 2016-04-06 | 2020-10-22 | Inserm (Institut National De La Santé Et La Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of age-related cardiometabolic diseases |
EP3443004A1 (en) | 2016-04-14 | 2019-02-20 | H. Hoffnabb-La Roche Ag | Anti-rspo3 antibodies and methods of use |
EP3443120A2 (en) | 2016-04-15 | 2019-02-20 | H. Hoffnabb-La Roche Ag | Methods for monitoring and treating cancer |
EP3443350B1 (en) | 2016-04-15 | 2020-12-09 | H. Hoffnabb-La Roche Ag | Methods for monitoring and treating cancer |
US20190125826A1 (en) | 2016-04-22 | 2019-05-02 | Inserm (Institut National De La Santé Et De La Médicale) | Methods and pharmaceutical composition for the treatment of inflammatory skin diseases associated with desmoglein-1 deficiency |
JP6871948B2 (en) | 2016-04-27 | 2021-05-19 | アッヴィ・インコーポレイテッド | Treatment of Diseases with Harmful IL-13 Activity Using Anti-IL-13 Antibodies |
AU2017259876A1 (en) | 2016-05-02 | 2018-10-25 | Ablynx Nv | Treatment of RSV infection |
UA123323C2 (en) | 2016-05-02 | 2021-03-17 | Ф. Хоффманн-Ля Рош Аг | The contorsbody - a single chain target binder |
US11098124B2 (en) | 2016-05-03 | 2021-08-24 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | CD31 shed as a molecular target for imaging of inflammation |
US11376269B2 (en) | 2016-05-06 | 2022-07-05 | Inserm | Pharmaceutical compositions for the treatment of chemoresistant acute myeloid leukemia (AML) |
WO2017194554A1 (en) | 2016-05-10 | 2017-11-16 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Combinations therapies for the treatment of cancer |
WO2017194441A1 (en) | 2016-05-11 | 2017-11-16 | F. Hoffmann-La Roche Ag | Modified anti-tenascin antibodies and methods of use |
EP3455254B1 (en) | 2016-05-11 | 2021-07-07 | F. Hoffmann-La Roche AG | Antigen binding molecules comprising a tnf family ligand trimer and a tenascin binding moiety |
EP3243836A1 (en) | 2016-05-11 | 2017-11-15 | F. Hoffmann-La Roche AG | C-terminally fused tnf family ligand trimer-containing antigen binding molecules |
HRP20221298T1 (en) | 2016-05-13 | 2022-12-23 | Bioatla, Inc. | Anti-ror2 antibodies, antibody fragments, their immunoconjugates and uses thereof |
EP3243832A1 (en) | 2016-05-13 | 2017-11-15 | F. Hoffmann-La Roche AG | Antigen binding molecules comprising a tnf family ligand trimer and pd1 binding moiety |
MA45029B1 (en) | 2016-05-18 | 2021-03-31 | Boehringer Ingelheim Int | Anti pd-1 and anti-lag3 antibodies for cancer treatment |
ES2858151T3 (en) | 2016-05-20 | 2021-09-29 | Hoffmann La Roche | PROTAC-Antibody Conjugates and Procedures for Use |
WO2017202962A1 (en) | 2016-05-24 | 2017-11-30 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of non small cell lung cancer (nsclc) that coexists with chronic obstructive pulmonary disease (copd) |
EP3465221B1 (en) | 2016-05-27 | 2020-07-22 | H. Hoffnabb-La Roche Ag | Bioanalytical method for the characterization of site-specific antibody-drug conjugates |
WO2017202890A1 (en) | 2016-05-27 | 2017-11-30 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for predicting and treating myeloma |
CN110603266A (en) | 2016-06-02 | 2019-12-20 | 豪夫迈·罗氏有限公司 | Type II anti-CD 20 and anti-CD 20/CD3 bispecific antibodies for the treatment of cancer |
EP3252078A1 (en) | 2016-06-02 | 2017-12-06 | F. Hoffmann-La Roche AG | Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer |
EP3464280B1 (en) | 2016-06-06 | 2021-10-06 | F. Hoffmann-La Roche AG | Silvestrol antibody-drug conjugates and methods of use |
CA3027047A1 (en) | 2016-06-08 | 2017-12-14 | Abbvie Inc. | Anti-cd98 antibodies and antibody drug conjugates |
AU2017277422A1 (en) | 2016-06-08 | 2019-01-03 | Abbvie Inc. | Anti-EGFR antibody drug conjugates |
CN109641962A (en) | 2016-06-08 | 2019-04-16 | 艾伯维公司 | Anti- B7-H3 antibody and antibody drug conjugates |
CN116059394A (en) | 2016-06-08 | 2023-05-05 | 艾伯维公司 | anti-EGFR antibody drug conjugates |
TW202304996A (en) | 2016-06-08 | 2023-02-01 | 美商艾伯維有限公司 | Anti-b7-h3 antibodies and antibody drug conjugates |
BR112018075653A2 (en) | 2016-06-08 | 2019-08-27 | Abbvie Inc | anti-b7-h3 antibodies and drug antibody conjugates |
WO2017214301A1 (en) | 2016-06-08 | 2017-12-14 | Abbvie Inc. | Anti-egfr antibody drug conjugates |
JP2019524651A (en) | 2016-06-08 | 2019-09-05 | アッヴィ・インコーポレイテッド | Anti-CD98 antibodies and antibody drug conjugates |
US20200147235A1 (en) | 2016-06-08 | 2020-05-14 | Abbvie Inc. | Anti-cd98 antibodies and antibody drug conjugates |
CN109475642B (en) | 2016-06-10 | 2023-05-02 | 卫材R&D管理有限公司 | Lysine conjugated immunoglobulins |
GB201610198D0 (en) | 2016-06-10 | 2016-07-27 | Ucb Biopharma Sprl | Anti-ige antibodies |
CN116143918A (en) | 2016-06-24 | 2023-05-23 | 豪夫迈·罗氏有限公司 | Anti-polyubiquitin multispecific antibodies |
WO2018007442A1 (en) | 2016-07-06 | 2018-01-11 | Ablynx N.V. | Treatment of il-6r related diseases |
HUE054228T2 (en) | 2016-07-15 | 2021-08-30 | Acceleron Pharma Inc | Compositions comprising actriia polypeptides for use in treating pulmonary hypertension |
CA3030926A1 (en) | 2016-07-19 | 2018-01-25 | Teva Pharmaceuticals Australia Pty Ltd. | Anti-cd47 combination therapy |
WO2018014260A1 (en) | 2016-07-20 | 2018-01-25 | Nanjing Legend Biotech Co., Ltd. | Multispecific antigen binding proteins and methods of use thereof |
BR112019001615A2 (en) | 2016-07-27 | 2019-04-30 | Acceleron Pharma Inc. | methods and compositions for treating myelofibrosis |
BR112019001693A2 (en) | 2016-07-29 | 2019-07-02 | Ct Hospitalier Universitaire Toulouse | antibodies targeting tumor-associated macrophages and their uses |
CN109415444B (en) | 2016-07-29 | 2024-03-01 | 中外制药株式会社 | Bispecific antibodies exhibiting increased functional activity of alternative FVIII cofactors |
CN109963871A (en) | 2016-08-05 | 2019-07-02 | 豪夫迈·罗氏有限公司 | Multivalence and multi-epitope Antibody and application method with agonist activity |
CN116271014A (en) | 2016-08-05 | 2023-06-23 | 中外制药株式会社 | Compositions for preventing or treating IL-8 related diseases |
JP7250674B2 (en) | 2016-08-08 | 2023-04-03 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | CANCER TREATMENT AND DIAGNOSTIC METHOD |
WO2018029182A1 (en) | 2016-08-08 | 2018-02-15 | Ablynx N.V. | Il-6r single variable domain antibodies for treatment of il-6r related diseases |
WO2018031662A1 (en) | 2016-08-11 | 2018-02-15 | Genentech, Inc. | Pyrrolobenzodiazepine prodrugs and antibody conjugates thereof |
WO2018041989A1 (en) | 2016-09-02 | 2018-03-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for diagnosing and treating refractory celiac disease type 2 |
EP3510407A1 (en) | 2016-09-08 | 2019-07-17 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for diagnosing and treating nephrotic syndrome |
US11098113B2 (en) | 2016-09-15 | 2021-08-24 | Vib Vzw | Immunoglobulin single variable domains directed against macrophage migration inhibitory factor |
SG10201607778XA (en) | 2016-09-16 | 2018-04-27 | Chugai Pharmaceutical Co Ltd | Anti-Dengue Virus Antibodies, Polypeptides Containing Variant Fc Regions, And Methods Of Use |
JOP20190009A1 (en) | 2016-09-21 | 2019-01-27 | Alx Oncology Inc | Antibodies against signal-regulatory protein alpha and methods of use |
SG11201900845YA (en) | 2016-09-23 | 2019-02-27 | Genentech Inc | Uses of il-13 antagonists for treating atopic dermatitis |
JP7051826B2 (en) | 2016-09-23 | 2022-04-11 | シーエスエル、リミテッド | Coagulation factor binding protein and its use |
MA46366A (en) | 2016-09-30 | 2019-08-07 | Janssen Biotech Inc | SAFE AND EFFECTIVE PROCESS FOR TREATING PSORIASIS WITH A SPECIFIC ANTIBODY AGAINST IL-23 |
WO2018060301A1 (en) | 2016-09-30 | 2018-04-05 | F. Hoffmann-La Roche Ag | Bispecific antibodies against cd3 |
CA3039573A1 (en) | 2016-10-05 | 2018-04-12 | Acceleron Pharma Inc. | Alk4:actriib heteromultimers and uses thereof |
CN116650622A (en) | 2016-10-05 | 2023-08-29 | 艾科赛扬制药股份有限公司 | Compositions and methods for treating kidney disease |
CN110139674B (en) | 2016-10-05 | 2023-05-16 | 豪夫迈·罗氏有限公司 | Method for preparing antibody drug conjugates |
WO2018068028A1 (en) | 2016-10-06 | 2018-04-12 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
WO2018068201A1 (en) | 2016-10-11 | 2018-04-19 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against ctla-4 |
WO2018075564A1 (en) | 2016-10-17 | 2018-04-26 | University Of Maryland, College Park | Multispecific antibodies targeting human immunodeficiency virus and methods of using the same |
EP3532496A1 (en) | 2016-10-28 | 2019-09-04 | Banyan Biomarkers, Inc. | Antibodies to ubiquitin c-terminal hydrolase l1 (uch-l1) and glial fibrillary acidic protein (gfap) and related methods |
CN110267678A (en) | 2016-10-29 | 2019-09-20 | 霍夫曼-拉罗奇有限公司 | Anti- MIC antibody and application method |
US20190345500A1 (en) | 2016-11-14 | 2019-11-14 | |Nserm (Institut National De La Santé Et De La Recherche Médicale) | Methods and pharmaceutical compositions for modulating stem cells proliferation or differentiation |
CN109923128A (en) | 2016-11-15 | 2019-06-21 | 基因泰克公司 | Administration for being treated with anti-CD20/ AntiCD3 McAb bispecific antibody |
WO2018093841A1 (en) | 2016-11-16 | 2018-05-24 | Janssen Biotech, Inc. | Method of treating psoriasis with anti-il-23 specific antibody |
JP7222888B2 (en) | 2016-11-16 | 2023-02-15 | アブリンクス エン.ヴェー. | T cell engaging polypeptides capable of binding CD123 and TCR alpha/beta |
EP3541830A1 (en) | 2016-11-17 | 2019-09-25 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for increasing endogenous protein level |
TW201829463A (en) | 2016-11-18 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | Anti-hla-g antibodies and use thereof |
JP7274417B2 (en) | 2016-11-23 | 2023-05-16 | イミュノア・セラピューティクス・インコーポレイテッド | 4-1BB binding protein and uses thereof |
TWI797097B (en) | 2016-11-28 | 2023-04-01 | 日商中外製藥股份有限公司 | Polypeptides comprising an antigen-binding domain and a transport moiety |
WO2018099968A1 (en) | 2016-11-29 | 2018-06-07 | Ablynx N.V. | Treatment of infection by respiratory syncytial virus (rsv) |
WO2018100190A1 (en) | 2016-12-02 | 2018-06-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for diagnosing renal cell carcinoma |
AU2017373889A1 (en) | 2016-12-07 | 2019-06-06 | Ac Immune Sa | Anti-Tau antibodies and methods of use |
CA3045294A1 (en) | 2016-12-07 | 2018-06-14 | Genentech, Inc. | Anti-tau antibodies and methods of use |
CN110114674B (en) | 2016-12-13 | 2023-05-09 | 豪夫迈·罗氏有限公司 | Method for determining the presence of a target antigen in a tumor sample |
US20190310250A1 (en) | 2016-12-16 | 2019-10-10 | Merck Patent Gmbh | Methods for the use of galectin 3 binding protein detected in the urine for monitoring the severity and progression of lupus nephritis |
IL267284B2 (en) | 2016-12-19 | 2023-03-01 | Hoffmann La Roche | Combination therapy with targeted 4-1bb (cd137) agonists |
JP7247091B2 (en) | 2016-12-20 | 2023-03-28 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with anti-CD20/anti-CD3 bispecific antibody and 4-1BB (CD137) agonist |
GB201621806D0 (en) | 2016-12-21 | 2017-02-01 | Philogen Spa | Immunocytokines with progressive activation mechanism |
EP3360898A1 (en) | 2017-02-14 | 2018-08-15 | Boehringer Ingelheim International GmbH | Bispecific anti-tnf-related apoptosis-inducing ligand receptor 2 and anti-cadherin 17 binding molecules for the treatment of cancer |
CN108261544B (en) | 2016-12-30 | 2023-05-05 | 江苏太平洋美诺克生物药业股份有限公司 | Stable pharmaceutical formulation comprising CD147 monoclonal antibody |
CN108261391B (en) | 2016-12-30 | 2022-03-01 | 江苏太平洋美诺克生物药业有限公司 | Stable pharmaceutical formulation comprising CD147 monoclonal antibody |
MX2019007795A (en) | 2017-01-03 | 2019-08-16 | Hoffmann La Roche | Bispecific antigen binding molecules comprising anti-4-1bb clone 20h4.9. |
WO2018134389A1 (en) | 2017-01-23 | 2018-07-26 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating infections |
CN110234351A (en) | 2017-01-30 | 2019-09-13 | 詹森生物科技公司 | For treating the anti-TNF antibodies, composition and method of activity psoriatic arthritis |
EP3576789A4 (en) | 2017-02-01 | 2020-11-25 | Centrymed Pharmaceuticals Inc. | MONOMERIC HUMAN IgG1 Fc AND BISPECIFIC ANTIBODIES |
MX2019009377A (en) | 2017-02-07 | 2019-12-11 | Janssen Biotech Inc | Anti-tnf antibodies, compositions, and methods for the treatment of active ankylosing spondylitis. |
US11266745B2 (en) | 2017-02-08 | 2022-03-08 | Imaginab, Inc. | Extension sequences for diabodies |
JP7341060B2 (en) | 2017-02-10 | 2023-09-08 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Methods and pharmaceutical compositions for the treatment of cancer associated with MAPK pathway activation |
MY197534A (en) | 2017-02-10 | 2023-06-21 | Genentech Inc | Anti-tryptase antibodies, compositions thereof, and uses thereof |
WO2018158335A1 (en) | 2017-02-28 | 2018-09-07 | Vib Vzw | Means and methods for oral protein delivery |
AU2018228873A1 (en) | 2017-03-01 | 2019-08-29 | Genentech, Inc. | Diagnostic and therapeutic methods for cancer |
EP3589654A1 (en) | 2017-03-02 | 2020-01-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antibodies having specificity to nectin-4 and uses thereof |
CR20190481A (en) | 2017-03-22 | 2020-01-06 | Genentech Inc | Optimized antibody compositions for treatment of ocular disorders |
MA49265A (en) | 2017-03-22 | 2020-02-05 | Ascendis Pharma As | Hydrogel cross-linked hyaluronic acid prodrug compositions and methods |
US20210186982A1 (en) | 2017-03-24 | 2021-06-24 | Universite Nice Sophia Antipolis | Methods and compositions for treating melanoma |
WO2018178029A1 (en) | 2017-03-27 | 2018-10-04 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating degenerative muscular and/or neurological conditions or diseases |
WO2018178030A1 (en) | 2017-03-27 | 2018-10-04 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating degenerative muscular and/or neurological conditions or diseases |
CA3056837A1 (en) | 2017-03-27 | 2018-10-04 | F. Hoffmann-La Roche Ag | Improved antigen binding receptors |
BR112019017629A2 (en) | 2017-03-27 | 2020-04-07 | Hoffmann La Roche | antigen-binding receptor, isolated polynucleotide, vector, transduced t cell, methods for treating a disease and for inducing lysis, use of the receptor and receptor |
TW202400231A (en) | 2017-03-28 | 2024-01-01 | 美商建南德克公司 | Methods of treating neurodegenerative diseases |
EP3601325B1 (en) | 2017-03-28 | 2023-07-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New tau species |
CN110573528B (en) | 2017-03-29 | 2023-06-09 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules to costimulatory TNF receptors |
EP3601346A1 (en) | 2017-03-29 | 2020-02-05 | H. Hoffnabb-La Roche Ag | Bispecific antigen binding molecule for a costimulatory tnf receptor |
WO2018178074A1 (en) | 2017-03-29 | 2018-10-04 | F. Hoffmann-La Roche Ag | Trimeric antigen binding molecules specific for a costimulatory tnf receptor |
US20200188541A1 (en) | 2017-03-30 | 2020-06-18 | Duke University | Radiolabeled biomolecules and their use |
JP7148539B2 (en) | 2017-04-03 | 2022-10-05 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | immunoconjugate |
EP3606963B1 (en) | 2017-04-03 | 2023-08-30 | F. Hoffmann-La Roche AG | Antibodies binding to steap-1 |
BR112019017329A2 (en) | 2017-04-03 | 2020-04-14 | Hoffmann La Roche | immunoconjugates, one or more polynucleotides and vectors, methods for the production of an immunoconjugate, treatment of a disease and for the stimulation of the immune system, composition, use of the immunoconjugate, invention and uses of the composition |
SG11201908787WA (en) | 2017-04-04 | 2019-10-30 | Hoffmann La Roche | Novel bispecific antigen binding molecules capable of specific binding to cd40 and to fap |
KR102408873B1 (en) | 2017-04-05 | 2022-06-15 | 에프. 호프만-라 로슈 아게 | Bispecific antibodies specifically binding to pd1 and lag3 |
KR102294136B1 (en) | 2017-04-05 | 2021-08-26 | 에프. 호프만-라 로슈 아게 | anti-LAG3 antibody |
CN108728444A (en) | 2017-04-18 | 2018-11-02 | 长春华普生物技术股份有限公司 | Immunoregulation polynucleotide and its application |
WO2018192974A1 (en) | 2017-04-18 | 2018-10-25 | Université Libre de Bruxelles | Biomarkers and targets for proliferative diseases |
WO2018195302A1 (en) | 2017-04-19 | 2018-10-25 | Bluefin Biomedicine, Inc. | Anti-vtcn1 antibodies and antibody drug conjugates |
MA49131A (en) | 2017-04-21 | 2020-03-25 | Hoffmann La Roche | USE OF KLK5 ANTAGONISTS FOR THE TREATMENT OF DISEASE |
TWI791519B (en) | 2017-04-27 | 2023-02-11 | 美商提薩羅有限公司 | Antibody agents directed against lymphocyte activation gene-3 (lag-3) and uses thereof |
JP2020518638A (en) | 2017-05-05 | 2020-06-25 | アラコス インコーポレイテッド | Methods and compositions for treating allergic eye diseases |
JP2020519261A (en) | 2017-05-11 | 2020-07-02 | ブイアイビー ブイゼットダブリュVib Vzw | Glycosylation of variable immunoglobulin domains |
WO2018213097A1 (en) | 2017-05-15 | 2018-11-22 | University Of Rochester | Broadly neutralizing anti-influenza monoclonal antibody and uses thereof |
EP3403649A1 (en) | 2017-05-16 | 2018-11-21 | Bayer Pharma Aktiengesellschaft | Inhibitors and antagonists of gpr84 for the treatment of endometriosis |
EP3624780A1 (en) | 2017-05-17 | 2020-03-25 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Flt3 inhibitors for improving pain treatments by opioids |
EP3406253A1 (en) | 2017-05-24 | 2018-11-28 | Bayer Aktiengesellschaft | Inhibitors and antagonists of human pycr1 |
WO2018218068A1 (en) * | 2017-05-24 | 2018-11-29 | Development Center For Biotechnology | Humanized antibodies against globo h and uses thereof in cancer treatments |
EP3409322A1 (en) | 2017-06-01 | 2018-12-05 | F. Hoffmann-La Roche AG | Treatment method |
EP4272822A3 (en) | 2017-06-02 | 2024-03-27 | Merck Patent GmbH | Adamts binding immunoglobulins |
WO2018220236A1 (en) | 2017-06-02 | 2018-12-06 | Merck Patent Gmbh | Polypeptides binding adamts5, mmp13 and aggrecan |
BR112019025392A2 (en) | 2017-06-02 | 2020-07-07 | Ablynx N.V. | aggrecan-binding immunoglobulins |
CA3064469A1 (en) | 2017-06-02 | 2018-12-06 | Merck Patent Gmbh | Mmp13 binding immunoglobulins |
EP3634582A1 (en) | 2017-06-08 | 2020-04-15 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating hyperpigmentation disorders |
EP3635398A1 (en) | 2017-06-08 | 2020-04-15 | Institut National de la Sante et de la Recherche Medicale (INSERM) | Chimeric receptor for use in whole-cell sensors for detecting analytes of interest |
TWI726217B (en) * | 2017-06-15 | 2021-05-01 | 財團法人生物技術開發中心 | Antibody-drug conjugates containing anti-globo h antibodies and uses thereof |
US20210403573A1 (en) | 2017-06-22 | 2021-12-30 | INSERM (Institut National de la Santé et de la Recherche Médicale | Methods and pharmaceutical compositions for the treatment of fibrosis with agents capable of inhibiting the activation of mucosal-associated invariant t (mait) cells |
WO2019000223A1 (en) | 2017-06-27 | 2019-01-03 | Nanjing Legend Biotech Co., Ltd. | Chimeric antibody immune effctor cell engagers and methods of use thereof |
WO2019002548A1 (en) | 2017-06-29 | 2019-01-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Treating migraine by agonising trek1, trek2 or heteromers including them |
TWI820031B (en) | 2017-07-11 | 2023-11-01 | 美商坎伯斯治療有限責任公司 | Agonist antibodies that bind human cd137 and uses thereof |
KR102625929B1 (en) | 2017-07-19 | 2024-01-16 | 브이아이비 브이지더블유 | Serum albumin binder |
WO2019016310A1 (en) | 2017-07-20 | 2019-01-24 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cancers |
TWI823859B (en) | 2017-07-21 | 2023-12-01 | 美商建南德克公司 | Therapeutic and diagnostic methods for cancer |
WO2019020480A1 (en) | 2017-07-24 | 2019-01-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antibodies and peptides to treat hcmv related diseases |
EP3658173A1 (en) | 2017-07-25 | 2020-06-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for modulating monocytopoiesis |
JP7407699B2 (en) | 2017-07-28 | 2024-01-04 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Bispecific antibody preparation |
JP7299160B2 (en) | 2017-08-03 | 2023-06-27 | アレクトル エルエルシー | ANTI-CD33 ANTIBODY AND METHOD OF USE THEREOF |
EP3444275A1 (en) | 2017-08-16 | 2019-02-20 | Exiris S.r.l. | Monoclonal antibody anti-fgfr4 |
CN111511762A (en) | 2017-08-21 | 2020-08-07 | 天演药业公司 | anti-CD137 molecules and uses thereof |
EP3676296A1 (en) | 2017-08-30 | 2020-07-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti-mesothelin radiolabelled single domain antibodies suitable for the imaging and treatment of cancers |
EP3457139A1 (en) | 2017-09-19 | 2019-03-20 | Promise Advanced Proteomics | Antibody-like peptides for quantifying therapeutic antibodies |
WO2019059411A1 (en) | 2017-09-20 | 2019-03-28 | Chugai Seiyaku Kabushiki Kaisha | Dosage regimen for combination therapy using pd-1 axis binding antagonists and gpc3 targeting agent |
EP3684471A1 (en) | 2017-09-20 | 2020-07-29 | Institut National de la Sante et de la Recherche Medicale (INSERM) | Methods and pharmaceutical compositions for modulating autophagy |
TW201922780A (en) | 2017-09-25 | 2019-06-16 | 美商健生生物科技公司 | Safe and effective method of treating Lupus with anti-IL12/IL23 antibody |
JP7450535B2 (en) | 2017-10-20 | 2024-03-15 | エフ. ホフマン-ラ ロシュ アーゲー | Method for generating multispecific antibodies from monospecific antibodies |
WO2019081456A1 (en) | 2017-10-24 | 2019-05-02 | Bayer Aktiengesellschaft | Use of activators and stimulators of sgc comprising a beta2 subunit |
AU2018358883A1 (en) | 2017-10-30 | 2020-04-23 | F. Hoffmann-La Roche Ag | Method for in vivo generation of multispecific antibodies from monospecific antibodies |
WO2019089753A2 (en) | 2017-10-31 | 2019-05-09 | Compass Therapeutics Llc | Cd137 antibodies and pd-1 antagonists and uses thereof |
WO2019086548A1 (en) | 2017-10-31 | 2019-05-09 | Vib Vzw | Novel antigen-binding chimeric proteins and methods and uses thereof |
JP2021500930A (en) | 2017-11-01 | 2021-01-14 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | COMP Body-Multivalent Target Binding Substance |
JP2021501162A (en) | 2017-11-01 | 2021-01-14 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with targeted OX40 agonist |
WO2019086499A1 (en) | 2017-11-01 | 2019-05-09 | F. Hoffmann-La Roche Ag | Novel tnf family ligand trimer-containing antigen binding molecules |
CR20200171A (en) | 2017-11-01 | 2020-06-14 | Hoffmann La Roche | Bispecific 2+1 contorsbodies |
TW201923089A (en) | 2017-11-06 | 2019-06-16 | 美商建南德克公司 | Diagnostic and therapeutic methods for cancer |
CA3082365A1 (en) | 2017-11-09 | 2019-05-16 | Pinteon Therapeutics Inc. | Methods and compositions for the generation and use of humanized conformation-specific phosphorylated tau antibodies |
EP3710486A1 (en) | 2017-11-15 | 2020-09-23 | Novo Nordisk A/S | Factor x binders enhancing fx activation |
US11851497B2 (en) | 2017-11-20 | 2023-12-26 | Compass Therapeutics Llc | CD137 antibodies and tumor antigen-targeting antibodies and uses thereof |
WO2019101995A1 (en) | 2017-11-27 | 2019-05-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for cardiac regeneration |
SG11202004897XA (en) | 2017-11-28 | 2020-06-29 | Chugai Pharmaceutical Co Ltd | Polypeptide including antigen-binding domain and carrying section |
US20210179721A1 (en) | 2017-11-29 | 2021-06-17 | Csl Limited | Method of treating or preventing ischemia-reperfusion injury |
BR112020010937A2 (en) | 2017-12-01 | 2020-11-17 | Seattle Genetics, Inc. | humanized anti-liv1 antibodies for the treatment of breast cancer |
CN111448213A (en) | 2017-12-01 | 2020-07-24 | 西雅图基因公司 | CD47 antibodies and their use for treating cancer |
TW201934578A (en) | 2017-12-14 | 2019-09-01 | 瑞士商赫孚孟拉羅股份公司 | treatment method |
EA202091521A1 (en) | 2017-12-19 | 2020-10-22 | Дзе Рокфеллер Юниверсити | HUMAN Fc IgG DOMAIN OPTIONS WITH IMPROVED EFFECTIVE FUNCTION |
EP3502140A1 (en) | 2017-12-21 | 2019-06-26 | F. Hoffmann-La Roche AG | Combination therapy of tumor targeted icos agonists with t-cell bispecific molecules |
CN111527107A (en) | 2017-12-21 | 2020-08-11 | 豪夫迈·罗氏有限公司 | Antibodies that bind HLA-A2/WT1 |
JP7394058B2 (en) | 2017-12-21 | 2023-12-07 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Universal reporter cell assay for specificity testing of novel antigen-binding moieties |
WO2019122060A1 (en) | 2017-12-21 | 2019-06-27 | F. Hoffmann-La Roche Ag | Car-t cell assay for specificity test of novel antigen binding moieties |
WO2019126472A1 (en) | 2017-12-22 | 2019-06-27 | Genentech, Inc. | Use of pilra binding agents for treatment of a disease |
TW201930358A (en) | 2017-12-28 | 2019-08-01 | 大陸商南京傳奇生物科技有限公司 | Single-domain antibodies and variants thereof against TIGIT |
JP2021508498A (en) | 2017-12-29 | 2021-03-11 | アレクター リミテッド ライアビリティ カンパニー | Anti-TMEM106B antibody and how to use it |
EP3735590A1 (en) | 2018-01-04 | 2020-11-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma resistant |
KR20200120641A (en) | 2018-01-15 | 2020-10-21 | 난징 레전드 바이오테크 씨오., 엘티디. | Single-domain antibody against PD-1 and variants thereof |
EP3740505A1 (en) | 2018-01-16 | 2020-11-25 | Lakepharma Inc. | Bispecific antibody that binds cd3 and another target |
EP3743096A1 (en) | 2018-01-25 | 2020-12-02 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antagonists of il-33 for use in methods for preventing ischemia reperfusion injury in an organ |
JP2021511793A (en) | 2018-01-31 | 2021-05-13 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Bispecific antibody containing an antigen binding site that binds to LAG3 |
EP3746476A1 (en) | 2018-01-31 | 2020-12-09 | Alector LLC | Anti-ms4a4a antibodies and methods of use thereof |
CN111630063A (en) | 2018-01-31 | 2020-09-04 | 豪夫迈·罗氏有限公司 | Stabilized immunoglobulin domains |
WO2019150309A1 (en) | 2018-02-02 | 2019-08-08 | Hammack Scott | Modulators of gpr68 and uses thereof for treating and preventing diseases |
WO2019148444A1 (en) | 2018-02-02 | 2019-08-08 | Adagene Inc. | Anti-ctla4 antibodies and methods of making and using the same |
KR20200118444A (en) | 2018-02-06 | 2020-10-15 | 아블린쓰 엔.브이. | Method of treatment of early episodes of TTP using immunoglobulin single variable domain |
CR20200391A (en) | 2018-02-08 | 2020-10-19 | Genentech Inc | Bispecific antigen-binding molecules and methods of use |
TWI829667B (en) | 2018-02-09 | 2024-01-21 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to gprc5d |
WO2019157358A1 (en) | 2018-02-09 | 2019-08-15 | Genentech, Inc. | Therapeutic and diagnostic methods for mast cell-mediated inflammatory diseases |
WO2019158675A1 (en) | 2018-02-16 | 2019-08-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating vitiligo |
TW202000702A (en) | 2018-02-26 | 2020-01-01 | 美商建南德克公司 | Dosing for treatment with anti-TIGIT and anti-PD-L1 antagonist antibodies |
EP3758742A1 (en) | 2018-03-01 | 2021-01-06 | Vrije Universiteit Brussel | Human pd-l1-binding immunoglobulins |
JP2021514648A (en) | 2018-03-01 | 2021-06-17 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Specificity assay for novel target antigen binding moieties |
KR20200129125A (en) | 2018-03-05 | 2020-11-17 | 얀센 바이오테크 인코포레이티드 | How to treat Crohn's disease with anti-IL23 specific antibodies |
TW202003561A (en) | 2018-03-13 | 2020-01-16 | 瑞士商赫孚孟拉羅股份公司 | Combination therapy with targeted 4-1BB (CD137) agonists |
EP3765489A1 (en) | 2018-03-13 | 2021-01-20 | F. Hoffmann-La Roche AG | Therapeutic combination of 4-1 bb agonists with anti-cd20 antibodies |
AU2019235033A1 (en) | 2018-03-14 | 2020-07-30 | Beijing Xuanyi Pharmasciences Co., Ltd. | Anti-claudin 18.2 antibodies |
US20200040103A1 (en) | 2018-03-14 | 2020-02-06 | Genentech, Inc. | Anti-klk5 antibodies and methods of use |
KR20200132938A (en) | 2018-03-15 | 2020-11-25 | 추가이 세이야쿠 가부시키가이샤 | Anti-dengue virus antibodies with cross-reactivity against Zika virus and methods of use |
WO2019179365A1 (en) | 2018-03-20 | 2019-09-26 | WuXi Biologics Ireland Limited | Novel anti-lag-3 antibody polypeptide |
AU2019240111A1 (en) | 2018-03-21 | 2020-09-17 | ALX Oncology Inc. | Antibodies against signal-regulatory protein alpha and methods of use |
CA3088676A1 (en) | 2018-03-23 | 2019-09-26 | Universite Libre De Bruxelles | Wnt signaling agonist molecules |
JP2021519073A (en) | 2018-03-29 | 2021-08-10 | ジェネンテック, インコーポレイテッド | Regulation of lactogenic activity in mammalian cells |
EP3778639A4 (en) | 2018-04-02 | 2021-06-09 | Mab-Venture Biopharm Co., Ltd. | Lymphocyte activation gene-3 (lag-3) binding antibody and use thereof |
WO2019192973A1 (en) | 2018-04-04 | 2019-10-10 | F. Hoffmann-La Roche Ag | Diagnostic assays to detect tumor antigens in cancer patients |
WO2019193375A1 (en) | 2018-04-04 | 2019-10-10 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of fzd7 inhibitors for the treatment of retinal neovascularization |
TW202011029A (en) | 2018-04-04 | 2020-03-16 | 美商建南德克公司 | Methods for detecting and quantifying FGF21 |
EP3775883A1 (en) | 2018-04-04 | 2021-02-17 | F. Hoffmann-La Roche AG | Diagnostic assays to detect tumor antigens in cancer patients |
EP3552631A1 (en) | 2018-04-10 | 2019-10-16 | Inatherys | Antibody-drug conjugates and their uses for the treatment of cancer |
MX2020010732A (en) | 2018-04-13 | 2020-11-09 | Hoffmann La Roche | Her2-targeting antigen binding molecules comprising 4-1bbl. |
AR115052A1 (en) | 2018-04-18 | 2020-11-25 | Hoffmann La Roche | MULTI-SPECIFIC ANTIBODIES AND THE USE OF THEM |
AR114789A1 (en) | 2018-04-18 | 2020-10-14 | Hoffmann La Roche | ANTI-HLA-G ANTIBODIES AND THE USE OF THEM |
WO2019207030A1 (en) | 2018-04-26 | 2019-10-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for predicting a response with an immune checkpoint inhibitor in a patient suffering from a lung cancer |
WO2019213384A1 (en) | 2018-05-03 | 2019-11-07 | University Of Rochester | Anti-influenza neuraminidase monoclonal antibodies and uses thereof |
CA3100007A1 (en) | 2018-05-14 | 2019-11-21 | Werewolf Therapeutics, Inc. | Activatable interleukin-2 polypeptides and methods of use thereof |
BR112020023167A2 (en) | 2018-05-14 | 2021-02-09 | Werewolf Therapeutics, Inc. | activatable cytokine polypeptides and methods of using these |
AU2019269066B2 (en) | 2018-05-18 | 2022-10-06 | F. Hoffmann-La Roche Ag | Targeted intracellular delivery of large nucleic acids |
EP3569618A1 (en) | 2018-05-19 | 2019-11-20 | Boehringer Ingelheim International GmbH | Antagonizing cd73 antibody |
EA202092825A1 (en) | 2018-05-25 | 2021-04-22 | ЭЛЕКТОР ЭлЭлСи | ANTI-SIRPA ANTIBODIES AND METHODS OF THEIR APPLICATION |
WO2019230868A1 (en) | 2018-05-30 | 2019-12-05 | 中外製薬株式会社 | Ligand-binding molecule containing single domain antibody |
JP7390315B2 (en) | 2018-06-01 | 2023-12-01 | エーザイ・アール・アンド・ディー・マネジメント株式会社 | Splicing modulator antibody-drug conjugates and methods of use thereof |
KR20210016406A (en) | 2018-06-01 | 2021-02-15 | 에자이 알앤드디 매니지먼트 가부시키가이샤 | How to use splicing modifier |
EP3801613A1 (en) | 2018-06-04 | 2021-04-14 | Bayer Aktiengesellschaft | Inhibitors of shp2 |
EP3805400A4 (en) | 2018-06-04 | 2022-06-29 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule showing changed half-life in cytoplasm |
TW202016151A (en) | 2018-06-09 | 2020-05-01 | 德商百靈佳殷格翰國際股份有限公司 | Multi-specific binding proteins for cancer treatment |
CA3103629A1 (en) | 2018-06-15 | 2019-12-19 | Flagship Pioneering Innovations V, Inc. | Increasing immune activity through modulation of postcellular signaling factors |
WO2019244107A1 (en) | 2018-06-21 | 2019-12-26 | Daiichi Sankyo Company, Limited | Compositions including cd3 antigen binding fragments and uses thereof |
KR20210024550A (en) | 2018-06-23 | 2021-03-05 | 제넨테크, 인크. | PD-1 axis binding antagonist, platinum agent, and method of treating lung cancer using topoisomerase II inhibitor |
EA202190138A1 (en) | 2018-06-29 | 2021-05-27 | ЭЛЕКТОР ЭлЭлСи | ANTI-SIRP-BETA1 ANTIBODIES AND METHODS OF THEIR USE |
EP3818083A2 (en) | 2018-07-03 | 2021-05-12 | Elstar Therapeutics, Inc. | Anti-tcr antibody molecules and uses thereof |
TW202035447A (en) | 2018-07-04 | 2020-10-01 | 瑞士商赫孚孟拉羅股份公司 | Novel bispecific agonistic 4-1bb antigen binding molecules |
WO2020008083A1 (en) | 2018-07-05 | 2020-01-09 | Consejo Superior De Investigaciones Científicas | Therapeutic target in chemokine receptors for the screening of compounds useful for the treatment of pathological processes involving chemokine signaling |
KR20230065382A (en) | 2018-07-13 | 2023-05-11 | 알렉터 엘엘씨 | Anti-sortilin antibodies and methods of use thereof |
MX2021000558A (en) | 2018-07-18 | 2021-04-13 | Genentech Inc | Methods of treating lung cancer with a pd-1 axis binding antagonist, an antimetabolite, and a platinum agent. |
EP3824295A4 (en) | 2018-07-18 | 2022-04-27 | Janssen Biotech, Inc. | Sustained response predictors after treatment with anti-il23 specific antibody |
BR112021000727A2 (en) | 2018-07-20 | 2021-04-13 | Surface Oncology, Inc. | ANTI-CD112R COMPOSITIONS AND METHODS |
JP7401166B2 (en) | 2018-08-01 | 2023-12-19 | イムチェック セラピューティクス エスエーエス | Anti-BTN3A antibodies and their use in the treatment of cancer or infectious disorders |
JPWO2020027330A1 (en) | 2018-08-03 | 2021-08-19 | 中外製薬株式会社 | Antigen-binding molecule containing two antigen-binding domains linked to each other |
SG11202100601TA (en) | 2018-08-08 | 2021-02-25 | Genentech Inc | Use of tryptophan derivatives and l-methionine for protein formulation |
AU2019318031A1 (en) | 2018-08-10 | 2021-02-25 | Chugai Seiyaku Kabushiki Kaisha | Anti-CD137 antigen-binding molecule and utilization thereof |
TW202021618A (en) | 2018-08-17 | 2020-06-16 | 美商23與我有限公司 | Anti-il1rap antibodies and methods of use thereof |
US11548938B2 (en) | 2018-08-21 | 2023-01-10 | Quidel Corporation | DbpA antibodies and uses thereof |
TW202016307A (en) | 2018-08-31 | 2020-05-01 | 美商阿列克特有限責任公司 | Anti-cd33 antibodies and methods of use thereof |
GB201814281D0 (en) | 2018-09-03 | 2018-10-17 | Femtogenix Ltd | Cytotoxic agents |
US20210278420A1 (en) | 2018-09-05 | 2021-09-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating asthma and allergic diseases |
GB2576914A (en) | 2018-09-06 | 2020-03-11 | Kymab Ltd | Antigen-binding molecules comprising unpaired variable domains produced in mammals |
US20220047701A1 (en) | 2018-09-10 | 2022-02-17 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Combination of her2/neu antibody with heme for treating cancer |
WO2020053147A1 (en) | 2018-09-10 | 2020-03-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of an inhibitor of ntsr1 activation or expression for preventing weight loss, muscle loss, and protein blood level decrease in subjects in need thereof |
WO2020053125A1 (en) | 2018-09-10 | 2020-03-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for the treatment of neurofibromatosis |
US20220023265A1 (en) | 2018-09-17 | 2022-01-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of inhibitors of phosphatase activity of soluble epoxide for the treatment of cardiometabolic diseases |
TW202023542A (en) | 2018-09-18 | 2020-07-01 | 瑞士商赫孚孟拉羅股份公司 | Use of a cathepsin s inhibitor against the formation of anti-drug antibodies |
US20220073638A1 (en) | 2018-09-19 | 2022-03-10 | INSERM (Institut National de la Santé et de la Recherche Médicale | Methods and pharmaceutical composition for the treatment of cancers resistant to immune checkpoint therapy |
MX2021003214A (en) | 2018-09-19 | 2021-05-12 | Genentech Inc | Therapeutic and diagnostic methods for bladder cancer. |
WO2020061349A1 (en) | 2018-09-21 | 2020-03-26 | Genentech, Inc. | Diagnostic methods for triple-negative breast cancer |
FI13575Y1 (en) | 2018-09-24 | 2024-03-26 | Janssen Biotech Inc | IL12/IL23 antibody to be used in a safe and efficient method for treating ulcerative colitis |
EP3856772A1 (en) | 2018-09-25 | 2021-08-04 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Use of antagonists of th17 cytokines for the treatment of bronchial remodeling in patients suffering from allergic asthma |
BR112021005907A2 (en) | 2018-09-27 | 2021-08-10 | Xilio Development, Inc. | masked cytokines, nucleic acid, vector, host cell, methods for producing a masked cytokine, for treating or preventing a neoplastic disease and for treating or preventing a neoplastic inflammatory or autoimmune disease, composition, pharmaceutical composition and kit |
WO2020070062A1 (en) | 2018-10-01 | 2020-04-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of tim-3 inhibitors for the treatment of exacerbations in patients suffering from severe asthma |
CN112654641A (en) | 2018-10-01 | 2021-04-13 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules with trivalent binding to CD40 |
MX2021003548A (en) | 2018-10-01 | 2021-05-27 | Hoffmann La Roche | Bispecific antigen binding molecules comprising anti-fap clone 212. |
EP3861022A1 (en) | 2018-10-04 | 2021-08-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of mucosal inflammatory diseases |
WO2020070288A1 (en) | 2018-10-05 | 2020-04-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and systems for controlling the agonistic properties of antibody variable domains by light |
EP3636657A1 (en) | 2018-10-08 | 2020-04-15 | Ablynx N.V. | Chromatography-free antibody purification method |
WO2020074937A1 (en) | 2018-10-09 | 2020-04-16 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of alpha-v-integrin (cd51) inhibitors for the treatment of cardiac fibrosis |
TW202028244A (en) | 2018-10-09 | 2020-08-01 | 美商建南德克公司 | Methods and systems for determining synapse formation |
WO2020081493A1 (en) | 2018-10-16 | 2020-04-23 | Molecular Templates, Inc. | Pd-l1 binding proteins |
WO2020079162A1 (en) | 2018-10-18 | 2020-04-23 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for inducing full ablation of hematopoiesis |
EP3867646A1 (en) | 2018-10-18 | 2021-08-25 | F. Hoffmann-La Roche AG | Diagnostic and therapeutic methods for sarcomatoid kidney cancer |
CA3115110A1 (en) | 2018-10-24 | 2020-04-30 | F. Hoffmann-La Roche Ag | Conjugated chemical inducers of degradation and methods of use |
CA3117051A1 (en) | 2018-11-05 | 2020-05-14 | Genentech, Inc. | Methods of producing two chain proteins in prokaryotic host cells |
JP2022507253A (en) | 2018-11-13 | 2022-01-18 | コンパス セラピューティクス リミテッド ライアビリティ カンパニー | Multispecific binding constructs for checkpoint molecules and their use |
GB201818477D0 (en) | 2018-11-13 | 2018-12-26 | Emstopa Ltd | Tissue plasminogen activator antibodies and method of use thereof |
EP3883961A1 (en) | 2018-11-20 | 2021-09-29 | Takeda Vaccines, Inc. | Novel anti-zika virus antibodies and uses thereof |
WO2020104479A1 (en) | 2018-11-20 | 2020-05-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cancers and resistant cancers with anti transferrin receptor 1 antibodies |
MA55149A (en) | 2018-11-20 | 2021-09-29 | Janssen Biotech Inc | SAFE AND EFFECTIVE PROCESS FOR TREATING PSORIASIS WITH A SPECIFIC ANTI-IL-23 ANTIBODY |
CA3119798A1 (en) | 2018-12-06 | 2020-06-11 | Genentech, Inc. | Combination therapy of diffuse large b-cell lymphoma comprising an anti-cd79b immunoconjugates, an alkylating agent and an anti-cd20 antibody |
WO2020115261A1 (en) | 2018-12-07 | 2020-06-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
WO2020123275A1 (en) | 2018-12-10 | 2020-06-18 | Genentech, Inc. | Photocrosslinking peptides for site specific conjugation to fc-containing proteins |
WO2020120592A1 (en) | 2018-12-12 | 2020-06-18 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for predicting and treating melanoma |
JP2022512401A (en) | 2018-12-13 | 2022-02-03 | エーザイ・アール・アンド・ディー・マネジメント株式会社 | Harboxy Diene Splicing Regulatory Antibodies-Drug Conjugates and Their Usage |
WO2020120644A1 (en) | 2018-12-13 | 2020-06-18 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New anti tau svqivykpv epitope single domain antibody |
JP2022514561A (en) | 2018-12-18 | 2022-02-14 | ヤンセン バイオテツク,インコーポレーテツド | A safe and effective way to treat lupus with anti-IL12 / IL23 antibodies |
AR117327A1 (en) | 2018-12-20 | 2021-07-28 | 23Andme Inc | ANTI-CD96 ANTIBODIES AND METHODS OF USE OF THEM |
BR112021011939A2 (en) | 2018-12-21 | 2021-09-14 | F. Hoffmann-La Roche Ag | CD3 BINDING ANTIBODY, CD3 AND TYRP-1 BINDING ANTIBODIES, ISOLATED POLYNUCLEOTIDE, HOST CELL, METHOD FOR PRODUCING A CD3 BINDING ANTIBODY, PHARMACEUTICAL COMPOSITION AND USES OF THE ANTIBODY |
WO2020127885A1 (en) | 2018-12-21 | 2020-06-25 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Compositions for treating cancers and resistant cancers |
EP3898683A1 (en) | 2018-12-21 | 2021-10-27 | F. Hoffmann-La Roche AG | Tumor-targeted superagonistic cd28 antigen binding molecules |
EP3898984A1 (en) | 2018-12-21 | 2021-10-27 | Genentech, Inc. | Methods of producing polypeptides using a cell line resistant to apoptosis |
BR112021010374A2 (en) | 2018-12-21 | 2021-08-24 | 23Andme, Inc. | Anti-il-36 antibodies and methods of using them |
UA128001C2 (en) | 2018-12-21 | 2024-03-06 | Ф. Хоффманн-Ля Рош Аг | Tumor-targeted agonistic cd28 antigen binding molecules |
MX2021007768A (en) | 2018-12-26 | 2021-08-24 | Xilio Dev Inc | Anti-ctla4 antibodies and methods of use thereof. |
EP3902830A1 (en) | 2018-12-30 | 2021-11-03 | F. Hoffmann-La Roche AG | Anti-rabbit cd19 antibodies and methods of use |
EP4059569A1 (en) | 2019-01-03 | 2022-09-21 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Methods and pharmaceutical compositions for enhancing cd8+ t cell-dependent immune responses in subjects suffering from cancer |
MX2021008434A (en) | 2019-01-14 | 2021-09-23 | Genentech Inc | Methods of treating cancer with a pd-1 axis binding antagonist and an rna vaccine. |
EP3911676A1 (en) | 2019-01-15 | 2021-11-24 | Janssen Biotech, Inc. | Anti-tnf antibody compositions and methods for the treatment of juvenile idiopathic arthritis |
CA3124515A1 (en) | 2019-01-23 | 2020-07-30 | Genentech, Inc. | Methods of producing multimeric proteins in eukaryotic host cells |
KR20210118878A (en) | 2019-01-23 | 2021-10-01 | 얀센 바이오테크 인코포레이티드 | Anti-TNF antibody composition for use in a method of treating psoriatic arthritis |
US20220089770A1 (en) | 2019-01-24 | 2022-03-24 | Chugai Seiyaku Kabushiki Kaisha | Novel cancer antigens and antibodies of said antigens |
JP2022518796A (en) | 2019-01-28 | 2022-03-16 | メイプル バイオテック エルエルシー | PSMP antagonist for use in the treatment of lung, kidney, or liver fibrosis disorders |
GB201901197D0 (en) | 2019-01-29 | 2019-03-20 | Femtogenix Ltd | G-A Crosslinking cytotoxic agents |
EP3921031A1 (en) | 2019-02-04 | 2021-12-15 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Methods and compositions for modulating blood-brain barrier |
EP3921443A1 (en) | 2019-02-08 | 2021-12-15 | F. Hoffmann-La Roche AG | Diagnostic and therapeutic methods for cancer |
CN113728107B (en) | 2019-02-18 | 2022-06-24 | Atb治疗公司 | Method for producing conjugate-toxin fusion proteins in plant cells or whole plants |
WO2020169707A1 (en) | 2019-02-21 | 2020-08-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Foxo1 inhibitor for use in the treatment of latent virus infection |
WO2020169698A1 (en) | 2019-02-21 | 2020-08-27 | F. Hoffmann-La Roche Ag | Sensitization of cancer cells to tnf by bet inhibition |
CN113710706A (en) | 2019-02-27 | 2021-11-26 | 豪夫迈·罗氏有限公司 | Administration for anti-TIGIT antibody and anti-CD 20 antibody or anti-CD 38 antibody treatment |
EP3930744A1 (en) | 2019-03-01 | 2022-01-05 | Allogene Therapeutics, Inc. | Dll3 targeting chimeric antigen receptors and binding agents |
US20220137054A1 (en) | 2019-03-05 | 2022-05-05 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New biomarkers and biotargets in renal cell carcinoma |
CA3126728A1 (en) | 2019-03-08 | 2020-09-17 | Genentech, Inc. | Methods for detecting and quantifying membrane-associated proteins on extracellular vesicles |
CA3133383A1 (en) | 2019-03-14 | 2020-09-17 | Janssen Biotech, Inc. | Methods for producing anti-tnf antibody compositions |
CN113840838A (en) | 2019-03-14 | 2021-12-24 | 詹森生物科技公司 | Methods of manufacture of compositions for the production of anti-TNF antibodies |
JP2022524074A (en) | 2019-03-14 | 2022-04-27 | ジェネンテック, インコーポレイテッド | Treatment of cancer with HER2xCD3 bispecific antibodies in combination with anti-HER2 MAB |
CA3133395A1 (en) | 2019-03-14 | 2020-09-17 | Janssen Biotech, Inc. | Manufacturing methods for producing anti-il12/il23 antibody compositions |
KR20210141998A (en) | 2019-03-14 | 2021-11-23 | 얀센 바이오테크 인코포레이티드 | Method of making anti-TNF antibody composition |
AU2020241428A1 (en) | 2019-03-15 | 2021-08-12 | Cartesian Therapeutics, Inc. | Anti-BCMA chimeric antigen receptors |
KR20210141583A (en) | 2019-03-18 | 2021-11-23 | 얀센 바이오테크 인코포레이티드 | Methods of Treating Psoriasis in Children Using Anti-IL-12/IL-23 Antibodies |
US20220153875A1 (en) | 2019-03-19 | 2022-05-19 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule containing antigen-binding domain of which binding activity to antigen is changed depending on mta, and library for obtaining said antigen-binding domain |
EP3947446A1 (en) | 2019-03-25 | 2022-02-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Treatment of taupathy disorders by targeting new tau species |
KR20210150623A (en) | 2019-03-29 | 2021-12-10 | 제넨테크, 인크. | Modulators of cell surface protein interactions and related methods and compositions |
US20220249511A1 (en) | 2019-03-29 | 2022-08-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for the treatment of keloid, hypertrophic scars and/or hyperpigmentation disorders |
EP3947737A2 (en) | 2019-04-02 | 2022-02-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods of predicting and preventing cancer in patients having premalignant lesions |
WO2020201442A1 (en) | 2019-04-03 | 2020-10-08 | Orega Biotech | Combination therapies based on pd1 and il-17b inhibitors |
US20220249458A1 (en) | 2019-04-04 | 2022-08-11 | Bayer Aktiengesellschaft | Agonists Of Adiponectin |
SG11202109901TA (en) | 2019-04-09 | 2021-10-28 | Hospital For Special Surgery | Protein binders for irhom2 |
WO2020208124A1 (en) | 2019-04-12 | 2020-10-15 | F. Hoffmann-La Roche Ag | Treatment of cancer using a cea cd3 bispecific antibody and a wnt signaling inhibitor |
CN113677403A (en) | 2019-04-12 | 2021-11-19 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules comprising lipocalin muteins |
BR112021020867A2 (en) | 2019-04-19 | 2022-01-04 | Genentech Inc | Antibodies, nucleic acid, vector, host cell, method of producing an antibody, immunoconjugate, pharmaceutical formulation, uses of the antibody, method of treating an individual with cancer, and method of reducing clearance |
KR20220004028A (en) | 2019-04-26 | 2022-01-11 | 알로젠 테라퓨틱스 인코포레이티드 | Methods for making allogeneic CAR T cells |
WO2020221796A1 (en) | 2019-04-30 | 2020-11-05 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
EP3962493A2 (en) | 2019-05-03 | 2022-03-09 | Flagship Pioneering Innovations V, Inc. | Methods of modulating immune activity/level of irf or sting or of treating cancer, comprising the administration of a sting modulator and/or purinergic receptor modulator or postcellular signaling factor |
EP3962947A2 (en) | 2019-05-03 | 2022-03-09 | F. Hoffmann-La Roche AG | Methods of treating cancer with an anti-pd-l1 antibody |
KR20220007136A (en) | 2019-05-14 | 2022-01-18 | 제넨테크, 인크. | Methods of Use of Anti-CD79b Immunoconjugates to Treat Follicular Lymphoma |
MX2021013766A (en) | 2019-05-14 | 2022-02-21 | Werewolf Therapeutics Inc | Separation moieties and methods and use thereof. |
KR20220012883A (en) | 2019-05-23 | 2022-02-04 | 얀센 바이오테크 인코포레이티드 | A method of treating inflammatory bowel disease with a combination therapy of IL-23 and an antibody against TNF alpha |
WO2020239945A1 (en) | 2019-05-28 | 2020-12-03 | Vib Vzw | Cancer treatment by targeting plexins in the immune compartment |
US20220228116A1 (en) | 2019-05-28 | 2022-07-21 | Vib Vzw | Cd8+ t-cells lacking plexins and their application in cancer treatment |
CA3142580A1 (en) | 2019-06-03 | 2020-12-10 | Janssen Biotech, Inc. | Anti-tnf antibodies, compositions, and methods for the treatment of active ankylosing spondylitis |
MX2021014885A (en) | 2019-06-03 | 2022-04-06 | Janssen Biotech Inc | Anti-tnf antibody compositions, and methods for the treatment of psoriatic arthritis. |
JPWO2020246563A1 (en) | 2019-06-05 | 2020-12-10 | ||
WO2020246567A1 (en) | 2019-06-05 | 2020-12-10 | 中外製薬株式会社 | Protease substrate, and polypeptide including protease cleavage sequence |
JPWO2020246617A1 (en) | 2019-06-07 | 2020-12-10 | ||
AU2020291527A1 (en) | 2019-06-11 | 2022-01-20 | Alector Llc | Anti-Sortilin antibodies for use in therapy |
JP2022537031A (en) | 2019-06-20 | 2022-08-23 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Conformational single domain antibodies against protease nexin-1 and uses thereof |
TW202115124A (en) | 2019-06-26 | 2021-04-16 | 瑞士商赫孚孟拉羅股份公司 | Novel antigen binding molecules binding to cea |
WO2020260326A1 (en) | 2019-06-27 | 2020-12-30 | F. Hoffmann-La Roche Ag | Novel icos antibodies and tumor-targeted antigen binding molecules comprising them |
WO2021001289A1 (en) | 2019-07-02 | 2021-01-07 | F. Hoffmann-La Roche Ag | Immunoconjugates comprising a mutant interleukin-2 and an anti-cd8 antibody |
AR119393A1 (en) | 2019-07-15 | 2021-12-15 | Hoffmann La Roche | ANTIBODIES THAT BIND NKG2D |
WO2021016233A1 (en) | 2019-07-22 | 2021-01-28 | Seagen Inc. | Humanized anti-liv1 antibodies for the treatment of cancer |
CR20220078A (en) | 2019-07-31 | 2022-06-24 | Alector Llc | Anti-ms4a4a antibodies and methods of use thereof |
EP4003526A2 (en) | 2019-07-31 | 2022-06-01 | F. Hoffmann-La Roche AG | Antibodies binding to gprc5d |
JP2022543551A (en) | 2019-07-31 | 2022-10-13 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Antibody that binds to GPRC5D |
US20220275105A1 (en) | 2019-08-02 | 2022-09-01 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Neutralizing granzyme b for providing cardioprotection in a subject who experienced a myocardial infarction |
WO2021028752A1 (en) | 2019-08-15 | 2021-02-18 | Janssen Biotech, Inc. | Anti-tfn antibodies for treating type i diabetes |
WO2021048292A1 (en) | 2019-09-11 | 2021-03-18 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
KR20220062304A (en) | 2019-09-12 | 2022-05-16 | 제넨테크, 인크. | Compositions and methods for treating lupus nephritis |
US11918649B2 (en) | 2019-09-18 | 2024-03-05 | Molecular Templates, Inc. | PD-L1-binding molecules comprising Shiga toxin a subunit scaffolds |
CA3150999A1 (en) | 2019-09-18 | 2021-03-25 | James Thomas Koerber | Anti-klk7 antibodies, anti-klk5 antibodies, multispecific anti-klk5/klk7 antibodies, and methods of use |
EP4031580A1 (en) | 2019-09-20 | 2022-07-27 | F. Hoffmann-La Roche AG | Dosing for anti-tryptase antibodies |
CN112625130B (en) | 2019-09-24 | 2023-08-29 | 财团法人工业技术研究院 | Anti-TIGIT antibodies and methods of use |
EP4048693A1 (en) | 2019-09-27 | 2022-08-31 | F. Hoffmann-La Roche AG | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
US20220281997A1 (en) | 2019-09-27 | 2022-09-08 | Nanjing GenScript Biotech Co., Ltd. | Anti-VHH Domain Antibodies and Use Thereof |
CN114450304B (en) | 2019-09-27 | 2023-12-12 | 国家医疗保健研究所 | anti-Mullera tube inhibiting substance antibodies and uses thereof |
WO2021058729A1 (en) | 2019-09-27 | 2021-04-01 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti-müllerian inhibiting substance type i receptor antibodies and uses thereof |
WO2021063968A1 (en) | 2019-09-30 | 2021-04-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Method and composition for diagnosing chronic obstructive pulmonary disease |
WO2021064009A1 (en) | 2019-09-30 | 2021-04-08 | Scirhom Gmbh | Protein binders to irhom2 epitopes |
TW202128756A (en) | 2019-10-02 | 2021-08-01 | 德商百靈佳殷格翰國際股份有限公司 | Multi-specific binding proteins for cancer treatment |
EP4037714A1 (en) | 2019-10-03 | 2022-08-10 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for modulating macrophages polarization |
US20220363776A1 (en) | 2019-10-04 | 2022-11-17 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of ovarian cancer, breast cancer or pancreatic cancer |
BR112022007216A2 (en) | 2019-10-18 | 2022-08-23 | Genentech Inc | METHODS FOR TREATMENT OF DIFFUSE LYMPHOMA, KIT AND IMMUNOCONJUGATE |
WO2021078359A1 (en) | 2019-10-21 | 2021-04-29 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of inhibitors of cubilin for the treatment of chronic kidney diseases |
WO2021089513A1 (en) | 2019-11-05 | 2021-05-14 | F. Hoffmann-La Roche Ag | Treatment of cancer using a hla-a2/wt1 x cd3 bispecific antibody and lenalidomide |
EP4055388A1 (en) | 2019-11-06 | 2022-09-14 | Genentech, Inc. | Diagnostic and therapeutic methods for treatment of hematologic cancers |
WO2021090062A1 (en) | 2019-11-07 | 2021-05-14 | Eisai R&D Management Co., Ltd. | Anti-mesothelin eribulin antibody-drug conjugates and methods of use |
US20220390449A1 (en) | 2019-11-12 | 2022-12-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New serological marker for the latent form of toxoplasmosis |
MX2022005904A (en) | 2019-11-15 | 2022-09-07 | Pliant Therapeutics Inc | Compositions and methods for activation of integrins. |
WO2021110796A1 (en) | 2019-12-04 | 2021-06-10 | Bayer Aktiengesellschaft | Inhibitors of shp2 |
US20230040928A1 (en) | 2019-12-09 | 2023-02-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antibodies having specificity to her4 and uses thereof |
GB201918279D0 (en) | 2019-12-12 | 2020-01-29 | Vib Vzw | Glycosylated single chain immunoglobulin domains |
IL293827A (en) | 2019-12-13 | 2022-08-01 | Alector Llc | Anti-mertk antibodies and methods of use thereof |
CN114867494B9 (en) | 2019-12-13 | 2024-01-12 | 基因泰克公司 | anti-LY 6G6D antibodies and methods of use |
US20230114107A1 (en) | 2019-12-17 | 2023-04-13 | Flagship Pioneering Innovations V, Inc. | Combination anti-cancer therapies with inducers of iron-dependent cellular disassembly |
MX2022007635A (en) | 2019-12-18 | 2022-07-19 | Hoffmann La Roche | Antibodies binding to hla-a2/mage-a4. |
US20210188971A1 (en) | 2019-12-19 | 2021-06-24 | Quidel Corporation | Monoclonal antibody fusions |
AU2020412609A1 (en) | 2019-12-23 | 2022-06-16 | Genentech, Inc. | Apolipoprotein L1-specific antibodies and methods of use |
JPWO2021132166A1 (en) | 2019-12-23 | 2021-07-01 | ||
PE20221585A1 (en) | 2019-12-27 | 2022-10-06 | Chugai Pharmaceutical Co Ltd | ANTI-ANTIGEN-4 ANTIBODY ASSOCIATED WITH THE CYTOTOXIC T LYMPHOCYTE (CTLA-4) AND USE THEREOF |
JP2023509952A (en) | 2020-01-09 | 2023-03-10 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Novel 4-1BBL trimer-containing antigen-binding molecule |
CN110818795B (en) | 2020-01-10 | 2020-04-24 | 上海复宏汉霖生物技术股份有限公司 | anti-TIGIT antibodies and methods of use |
EP4090770A1 (en) | 2020-01-17 | 2022-11-23 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
WO2022050954A1 (en) | 2020-09-04 | 2022-03-10 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
WO2021194481A1 (en) | 2020-03-24 | 2021-09-30 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
IL294859A (en) | 2020-01-31 | 2022-09-01 | Genentech Inc | Methods of inducing neoepitope-specific t cells with a pd-1 axis binding antagonist and an rna vaccine |
CA3167027A1 (en) | 2020-02-05 | 2021-08-12 | Larimar Therapeutics, Inc. | Tat peptide binding proteins and uses thereof |
CA3167349A1 (en) | 2020-02-10 | 2021-08-19 | Qing Zhou | Claudin 18.2 antibody and use thereof |
KR20220139357A (en) | 2020-02-10 | 2022-10-14 | 상하이 에스쿠겐 바이오테크놀로지 컴퍼니 리미티드 | CLDN18.2 Antibodies and Their Uses |
TW202144395A (en) | 2020-02-12 | 2021-12-01 | 日商中外製藥股份有限公司 | Anti-CD137 antigen-binding molecule for use in cancer treatment |
CN113248611A (en) | 2020-02-13 | 2021-08-13 | 湖南华康恒健生物技术有限公司 | anti-BCMA antibody, pharmaceutical composition and application thereof |
WO2021170750A1 (en) | 2020-02-28 | 2021-09-02 | Orega Biotech | Combination therapies based on ctla4 and il-17b inhibitors |
CA3174680A1 (en) | 2020-03-13 | 2021-09-16 | Genentech, Inc. | Anti-interleukin-33 antibodies and uses thereof |
WO2021188749A1 (en) | 2020-03-19 | 2021-09-23 | Genentech, Inc. | Isoform-selective anti-tgf-beta antibodies and methods of use |
PE20230414A1 (en) | 2020-03-24 | 2023-03-07 | Genentech Inc | TIE2 FIXING AGENTS AND METHODS OF USE |
CA3169908A1 (en) | 2020-03-26 | 2021-09-30 | Genentech, Inc. | Modified mammalian cells having reduced host cell proteins |
WO2021202590A1 (en) | 2020-03-31 | 2021-10-07 | Alector Llc | Anti-mertk antibodies and methods of use thereof |
US20230121511A1 (en) | 2020-03-31 | 2023-04-20 | Chugai Seiyaku Kabushiki Kaisha | Method for producing multispecific antigen-binding molecules |
MX2022012376A (en) | 2020-03-31 | 2023-02-15 | Biotalys NV | Anti-fungal polypeptides. |
JP2023519930A (en) | 2020-04-01 | 2023-05-15 | ユニバーシティ オブ ロチェスター | Monoclonal Antibodies Against Hemagglutinin (HA) and Neuraminidase (NA) of Influenza H3N2 Virus |
AR121706A1 (en) | 2020-04-01 | 2022-06-29 | Hoffmann La Roche | OX40 AND FAP-TARGETED BSPECIFIC ANTIGEN-BINDING MOLECULES |
WO2021198511A1 (en) | 2020-04-03 | 2021-10-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treatment of sars-cov-2 infection |
JP2023520515A (en) | 2020-04-03 | 2023-05-17 | ジェネンテック, インコーポレイテッド | Therapeutic and diagnostic methods for cancer |
AU2021256936A1 (en) | 2020-04-15 | 2022-07-21 | F. Hoffmann-La Roche Ag | Immunoconjugates |
CN113527488A (en) | 2020-04-22 | 2021-10-22 | 迈威(上海)生物科技股份有限公司 | Single variable domain antibody targeting human programmed death ligand 1(PD-L1) and derivative thereof |
IL297541A (en) | 2020-04-24 | 2022-12-01 | Genentech Inc | Methods of using anti-cd79b immunoconjugates |
AR121918A1 (en) | 2020-04-24 | 2022-07-20 | Hoffmann La Roche | MODULATION OF ENZYMES AND PATHWAYS WITH SULFHYDRYL COMPOUNDS AND THEIR DERIVATIVES |
TW202206100A (en) | 2020-04-27 | 2022-02-16 | 美商西健公司 | Treatment for cancer |
MX2021015024A (en) | 2020-04-28 | 2022-01-18 | Univ Rockefeller | Neutralizing anti-sars-cov-2 antibodies and methods of use thereof. |
WO2021222167A1 (en) | 2020-04-28 | 2021-11-04 | Genentech, Inc. | Methods and compositions for non-small cell lung cancer immunotherapy |
IL297830A (en) | 2020-05-03 | 2023-01-01 | Levena Suzhou Biopharma Co Ltd | Antibody-drug conjugates (adcs) comprising an anti-trop-2 antibody, compositions comprising such adcs, as well as methods of making and using the same |
WO2021224401A1 (en) | 2020-05-07 | 2021-11-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for determining a reference range of β-galactose exposure platelet |
EP4149558A1 (en) | 2020-05-12 | 2023-03-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New method to treat cutaneous t-cell lymphomas and tfh derived lymphomas |
EP4149964A2 (en) | 2020-05-15 | 2023-03-22 | Apogenix AG | Multi-specific immune modulators |
WO2021229104A1 (en) | 2020-05-15 | 2021-11-18 | Université de Liège | Anti-cd38 single-domain antibodies in disease monitoring and treatment |
AU2021276332A1 (en) | 2020-05-19 | 2022-11-17 | Boehringer Ingelheim International Gmbh | Binding molecules for the treatment of cancer |
US20230302050A1 (en) | 2020-05-20 | 2023-09-28 | Institut Curie | Single Domain Antibodies and Their Use in Cancer Therapies |
US20230204567A1 (en) | 2020-05-20 | 2023-06-29 | Takeda Vaccines, Inc. | Method for determining the potency of antigens |
WO2021236845A1 (en) | 2020-05-20 | 2021-11-25 | Takeda Vaccines, Inc. | Method for detection of zika virus specific antibodies |
WO2021236225A1 (en) | 2020-05-20 | 2021-11-25 | Takeda Vaccines, Inc. | Method for detection of zika virus specific antibodies |
JP2023526477A (en) | 2020-05-20 | 2023-06-21 | アンスティテュ・クリー | Synthetic single domain library |
CN116323665A (en) | 2020-05-29 | 2023-06-23 | 23和我公司 | anti-CD 200R1 antibodies and methods of use thereof |
WO2021245224A1 (en) | 2020-06-05 | 2021-12-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for treating ocular diseases |
JP2023528412A (en) | 2020-06-05 | 2023-07-04 | エーザイ・アール・アンド・ディー・マネジメント株式会社 | Anti-BCMA antibody-drug conjugates and methods of use |
BR112022024996A2 (en) | 2020-06-08 | 2022-12-27 | Hoffmann La Roche | ANTIBODIES, NUCLEIC ACID, HOST CELL, METHOD FOR PRODUCING AN ANTIBODY, PHARMACEUTICAL COMPOSITION, THERAPEUTIC AGENT, USE OF THE ANTIBODY, AND METHOD FOR TREATING AN INDIVIDUAL WITH HEPATITIS B |
MX2022015651A (en) | 2020-06-11 | 2023-01-16 | Genentech Inc | Nanolipoprotein-polypeptide conjugates and compositions, systems, and methods using same. |
WO2021252977A1 (en) | 2020-06-12 | 2021-12-16 | Genentech, Inc. | Methods and compositions for cancer immunotherapy |
KR20230025691A (en) | 2020-06-16 | 2023-02-22 | 제넨테크, 인크. | Methods and compositions for treating triple negative breast cancer |
CA3181672A1 (en) | 2020-06-18 | 2021-12-23 | Shi Li | Treatment with anti-tigit antibodies and pd-1 axis binding antagonists |
TW202216767A (en) | 2020-06-19 | 2022-05-01 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to cd3 and folr1 |
KR20230025667A (en) | 2020-06-19 | 2023-02-22 | 에프. 호프만-라 로슈 아게 | Protease Activated T Cell Bispecific Antibody |
WO2021255146A1 (en) | 2020-06-19 | 2021-12-23 | F. Hoffmann-La Roche Ag | Antibodies binding to cd3 and cea |
BR112022025574A2 (en) | 2020-06-19 | 2023-01-03 | Hoffmann La Roche | ANTIBODIES THAT BINDS CD3, POLYNUCLEOTIDE ISOLATED, HOST CELL, METHOD FOR PRODUCING AN ANTIBODY THAT BINDS CD3 AND FOR TREAT A DISEASE IN AN INDIVIDUAL, PHARMACEUTICAL COMPOSITION, ANTIBODY FOR USE AND INVENTION |
CR20220637A (en) | 2020-06-19 | 2023-01-31 | Hoffmann La Roche | Antibodies binding to cd3 and cd19 |
CA3183475A1 (en) | 2020-06-22 | 2021-12-30 | Thomas Huber | Anti-il-36 antibodies and methods of use thereof |
TW202216769A (en) | 2020-06-23 | 2022-05-01 | 瑞士商赫孚孟拉羅股份公司 | Agonistic cd28 antigen binding molecules targeting her2 |
IL299161A (en) | 2020-06-24 | 2023-02-01 | Genentech Inc | Apoptosis resistant cell lines |
WO2021260064A1 (en) | 2020-06-25 | 2021-12-30 | F. Hoffmann-La Roche Ag | Anti-cd3/anti-cd28 bispecific antigen binding molecules |
JP2023532726A (en) | 2020-06-29 | 2023-07-31 | インサーム(インスティテュ ナシオナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシェ メディカル) | ANTI-PROTEIN S SINGLE DOMAIN ANTIBODY AND POLYPEPTIDE CONTAINING THE SAME |
EP4172323A1 (en) | 2020-06-29 | 2023-05-03 | Flagship Pioneering Innovations V, Inc. | Viruses engineered to promote thanotransmission and their use in treating cancer |
WO2022008597A1 (en) | 2020-07-08 | 2022-01-13 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of infectious diseases |
TW202204895A (en) | 2020-07-13 | 2022-02-01 | 美商建南德克公司 | Cell-based methods for predicting polypeptide immunogenicity |
BR112023000839A2 (en) | 2020-07-17 | 2023-02-07 | Genentech Inc | ISOLATED ANTIBODIES, ISOLATED NUCLEIC ACID, HOST CELL, METHODS FOR PRODUCING AN ANTIBODY THAT BINDS TO HUMAN NOTCH2, FOR TREATING AN INDIVIDUAL WITH A MUCO-OBSTRUCTIVE PULMONARY DISEASE, AND FOR REDUCING THE NUMBER OF SECRETORY CELLS IN AN INDIVIDUAL, PHARMACEUTICAL COMPOSITION, ANTIBODY, ANTIBODY FOR USE AND USE OF THE ANTIBODY |
MX2023000888A (en) | 2020-07-21 | 2023-02-22 | Genentech Inc | Antibody-conjugated chemical inducers of degradation of brm and methods thereof. |
GB2597532A (en) | 2020-07-28 | 2022-02-02 | Femtogenix Ltd | Cytotoxic compounds |
WO2022023379A1 (en) | 2020-07-28 | 2022-02-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for preventing and treating a cancer |
EP4189071A1 (en) | 2020-08-03 | 2023-06-07 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Population of treg cells functionally committed to exert a regulatory activity and their use for adoptive therapy |
EP4192942A1 (en) | 2020-08-07 | 2023-06-14 | Genentech, Inc. | T cell-based methods for predicting polypeptide immunogenicity |
IL300701A (en) | 2020-08-17 | 2023-04-01 | Atb Therapeutics | Recombinant immunotoxin comprising a ribotoxin or rnase |
CN114106173A (en) | 2020-08-26 | 2022-03-01 | 上海泰槿生物技术有限公司 | anti-OX 40 antibodies, pharmaceutical compositions and uses thereof |
WO2022047222A2 (en) | 2020-08-28 | 2022-03-03 | Genentech, Inc. | Crispr/cas9 multiplex knockout of host cell proteins |
CN116113707A (en) | 2020-08-31 | 2023-05-12 | 基因泰克公司 | Methods for producing antibodies |
JP2023541627A (en) | 2020-09-14 | 2023-10-03 | イシュノス サイエンシズ ソシエテ アノニム | Antibodies that bind to IL1RAP and uses thereof |
IL300543A (en) | 2020-09-24 | 2023-04-01 | Hoffmann La Roche | Prevention or mitigation of t-cell bispecific antibody-related adverse effects |
WO2022063947A1 (en) | 2020-09-24 | 2022-03-31 | Vib Vzw | Combination of p2y6 inhibitors and immune checkpoint inhibitors |
WO2022063957A1 (en) | 2020-09-24 | 2022-03-31 | Vib Vzw | Biomarker for anti-tumor therapy |
WO2022064049A1 (en) | 2020-09-28 | 2022-03-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Method for diagnosing brucella infection |
WO2022067348A1 (en) | 2020-09-28 | 2022-03-31 | Seagen Inc. | Humanized anti-liv1 antibodies for the treatment of cancer |
TW202229348A (en) | 2020-09-30 | 2022-08-01 | 美商西健公司 | Uveal melanoma treatment using sea-cd40 |
JP2023544407A (en) | 2020-10-05 | 2023-10-23 | ジェネンテック, インコーポレイテッド | Administration for treatment with anti-FcRH5/anti-CD3 bispecific antibodies |
EP4226155A1 (en) | 2020-10-09 | 2023-08-16 | Takeda Vaccines, Inc. | Methods for determining complement-fixing antibodies |
EP4229090A1 (en) | 2020-10-16 | 2023-08-23 | Université d'Aix-Marseille | Anti-gpc4 single domain antibodies |
AR123855A1 (en) | 2020-10-20 | 2023-01-18 | Genentech Inc | PEG-CONJUGATED ANTI-MERTK ANTIBODIES AND METHODS OF USE |
WO2022084300A1 (en) | 2020-10-20 | 2022-04-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for diagnosis and monitoring form of coronavirus infection |
EP4232040A1 (en) | 2020-10-20 | 2023-08-30 | F. Hoffmann-La Roche AG | Combination therapy of pd-1 axis binding antagonists and lrrk2 inhitibors |
WO2022084355A2 (en) | 2020-10-21 | 2022-04-28 | Boehringer Ingelheim International Gmbh | Agonistic trkb binding molecules for the treatment of eye diseases |
WO2022084531A1 (en) | 2020-10-23 | 2022-04-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating glioma |
WO2022093981A1 (en) | 2020-10-28 | 2022-05-05 | Genentech, Inc. | Combination therapy comprising ptpn22 inhibitors and pd-l1 binding antagonists |
JP2023549062A (en) | 2020-10-30 | 2023-11-22 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Treatment of cancer using CEA CD3 bispecific antibodies and TGFβ signaling inhibitors |
KR20230095119A (en) | 2020-11-04 | 2023-06-28 | 제넨테크, 인크. | Dosing for Treatment with Anti-CD20/Anti-CD3 Bispecific Antibodies |
EP4240493A2 (en) | 2020-11-04 | 2023-09-13 | Genentech, Inc. | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies and anti-cd79b antibody drug conjugates |
EP4240758A1 (en) | 2020-11-04 | 2023-09-13 | The Rockefeller University | Neutralizing anti-sars-cov-2 antibodies |
AU2021376354A1 (en) | 2020-11-04 | 2023-06-22 | Myeloid Therapeutics, Inc. | Engineered chimeric fusion protein compositions and methods of use thereof |
CA3196076A1 (en) | 2020-11-04 | 2022-05-12 | Chi-Chung Li | Subcutaneous dosing of anti-cd20/anti-cd3 bispecific antibodies |
EP4240761A1 (en) | 2020-11-05 | 2023-09-13 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Use of il-6 inhibitors for the treatment of acute chest syndrome in patients suffering from sickle cell disease |
EP4243856A1 (en) | 2020-11-10 | 2023-09-20 | F. Hoffmann-La Roche AG | Prevention or mitigation of t-cell engaging agent-related adverse effects |
WO2022101088A1 (en) | 2020-11-16 | 2022-05-19 | F. Hoffmann-La Roche Ag | Fab high mannose glycoforms |
WO2022101481A1 (en) | 2020-11-16 | 2022-05-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for predicting and treating uveal melanoma |
WO2022101458A1 (en) | 2020-11-16 | 2022-05-19 | F. Hoffmann-La Roche Ag | Combination therapy with fap-targeted cd40 agonists |
BR112023009875A2 (en) | 2020-11-23 | 2024-02-06 | Genentech Inc | METHODS TO TREAT AN INDIVIDUAL WITH A SARS-COV-2 INFECTION, TO REDUCE THE BINDING OF SARS-COV-2 TO A CELL, TO DECREASE THE SARS-COV-2 INFECTION, AND TO IDENTIFY A MODULATOR, METHODS OF PROPHYLAXIS, MODULATORS ISOLATES, USES AND ANTAGONISTS |
AU2021386240A1 (en) | 2020-11-27 | 2023-06-29 | Genentech, Inc. | Methods for modulating host cell surface interactions with human cytomegalovirus |
KR20230116843A (en) | 2020-12-01 | 2023-08-04 | 제넨테크, 인크. | Biological vesicles presenting cell surface proteins and methods associated therewith |
AU2021392039A1 (en) | 2020-12-02 | 2023-06-29 | Alector Llc | Methods of use of anti-sortilin antibodies |
US20220213199A1 (en) | 2020-12-17 | 2022-07-07 | Hoffmann-La Roche Inc. | Anti-HLA-G antibodies and use thereof |
CA3202897A1 (en) | 2020-12-21 | 2022-06-30 | Peter Joseph Gough | Use of cell turnover factors for increasing tissue regeneration |
IL304067A (en) | 2021-01-06 | 2023-08-01 | Hoffmann La Roche | Combination therapy employing a pd1-lag3 bispecific antibody and a cd20 t cell bispecific antibody |
WO2022153212A1 (en) | 2021-01-13 | 2022-07-21 | Axon Neuroscience Se | Antibodies neutralizing sars-cov-2 |
CA3204731A1 (en) | 2021-01-13 | 2022-07-21 | John T. POIRIER | Anti-dll3 antibody-drug conjugate |
EP4277664A1 (en) | 2021-01-13 | 2023-11-22 | Memorial Sloan Kettering Cancer Center | Antibody-pyrrolobenzodiazepine derivative conjugate |
JP2024505428A (en) | 2021-01-14 | 2024-02-06 | アンスティテュ キュリー | HER2 single domain antibody variants and their CARs |
WO2022155324A1 (en) | 2021-01-15 | 2022-07-21 | The Rockefeller University | Neutralizing anti-sars-cov-2 antibodies |
AR124681A1 (en) | 2021-01-20 | 2023-04-26 | Abbvie Inc | ANTI-EGFR ANTIBODY-DRUG CONJUGATES |
EP4281484A1 (en) | 2021-01-22 | 2023-11-29 | Bionecure Therapeutics, Inc. | Anti-her-2/trop-2 constructs and uses thereof |
KR20230137295A (en) | 2021-01-28 | 2023-10-04 | 난징 챔피언 바이오테크놀로지 컴퍼니 리미티드 | Conjugates and their uses |
CN117241804A (en) | 2021-02-17 | 2023-12-15 | 非营利性组织佛兰芒综合大学生物技术研究所 | Inhibition of SLC4A4 in cancer treatment |
CN117321076A (en) | 2021-02-19 | 2023-12-29 | 美国卫生及公众服务部代表 | Single domain antibodies neutralizing SARS-CoV-2 |
CA3207134A1 (en) | 2021-02-19 | 2022-08-25 | Jeffrey A. Ledbetter | Dnase fusion polypeptides and related compositions and methods |
WO2022178415A1 (en) | 2021-02-22 | 2022-08-25 | Genentech, Inc. | Methods for modulating host cell surface interactions with herpesviruses |
KR20230150287A (en) | 2021-02-26 | 2023-10-30 | 바이엘 악티엔게젤샤프트 | Inhibitors of IL-11 or IL-11RA for use in the treatment of abnormal uterine bleeding |
US20220306743A1 (en) | 2021-03-01 | 2022-09-29 | Xilio Development, Inc. | Combination of ctla4 and pd1/pdl1 antibodies for treating cancer |
BR112023017035A2 (en) | 2021-03-01 | 2024-02-06 | New York Soc For The Relief Of The Ruptured And Crippled Maintaining The Hospital For Special Surger | HUMANIZED ANTIBODIES AGAINST IRHOM2, NUCLEIC ACID, USE OF THE ANTIBODY, PHARMACEUTICAL COMPOSITION, COMPOSITION, TREATMENT METHOD AND THERAPEUTIC KIT |
TW202246324A (en) | 2021-03-01 | 2022-12-01 | 美商艾希利歐發展股份有限公司 | Combination of masked ctla4 and pd1/pdl1 antibodies for treating cancer |
EP4301418A1 (en) | 2021-03-03 | 2024-01-10 | Sorrento Therapeutics, Inc. | Antibody-drug conjugates comprising an anti-bcma antibody |
WO2022190034A1 (en) | 2021-03-12 | 2022-09-15 | Janssen Biotech, Inc. | Method of treating psoriatic arthritis patients with inadequate response to tnf therapy with anti-il23 specific antibody |
JP2024512377A (en) | 2021-03-12 | 2024-03-19 | ジェネンテック, インコーポレイテッド | Anti-KLK7 antibodies, anti-KLK5 antibodies, multispecific anti-KLK5/KLK7 antibodies, and methods of use |
US20220298236A1 (en) | 2021-03-12 | 2022-09-22 | Janssen Biotech, Inc. | Safe and Effective Method of Treating Psoriatic Arthritis with Anti-IL23 Specific Antibody |
WO2022198192A1 (en) | 2021-03-15 | 2022-09-22 | Genentech, Inc. | Compositions and methods of treating lupus nephritis |
WO2022197945A1 (en) | 2021-03-17 | 2022-09-22 | Molecular Templates, Inc. | Pd-l1 binding proteins comprising shiga toxin a subunit scaffolds and cd8+ t cell antigens |
WO2022194908A1 (en) | 2021-03-17 | 2022-09-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
CA3208011A1 (en) | 2021-03-17 | 2022-09-22 | Sarah Harris | Methods of treating atopic dermatitis with anti il-13 antibodies |
JP2024512002A (en) | 2021-03-18 | 2024-03-18 | アレクトル エルエルシー | Anti-TMEM106B antibody and method of use thereof |
WO2022197877A1 (en) | 2021-03-19 | 2022-09-22 | Genentech, Inc. | Methods and compositions for time delayed bio-orthogonal release of cytotoxic agents |
EP4313317A1 (en) | 2021-03-23 | 2024-02-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for the diagnosis and treatment of t cell-lymphomas |
JP2024511610A (en) | 2021-03-23 | 2024-03-14 | アレクトル エルエルシー | Anti-TMEM106B antibody for treatment and prevention of coronavirus infection |
WO2022207652A1 (en) | 2021-03-29 | 2022-10-06 | Scirhom Gmbh | Methods of treatment using protein binders to irhom2 epitopes |
CN117157312A (en) | 2021-03-30 | 2023-12-01 | 豪夫迈·罗氏有限公司 | Protease-activated polypeptides |
EP4313109A1 (en) | 2021-03-31 | 2024-02-07 | Flagship Pioneering Innovations V, Inc. | Thanotransmission polypeptides and their use in treating cancer |
BR112023020832A2 (en) | 2021-04-08 | 2023-12-19 | Marengo Therapeutics Inc | TCR-BINDED MULTIFUNCTIONAL MOLECULES AND THEIR USES |
WO2022214681A1 (en) | 2021-04-09 | 2022-10-13 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for the treatment of anaplastic large cell lymphoma |
EP4323402A1 (en) | 2021-04-14 | 2024-02-21 | Villaris Therapeutics, Inc. | Anti-cd122 antibodies and uses thereof |
AR125344A1 (en) | 2021-04-15 | 2023-07-05 | Chugai Pharmaceutical Co Ltd | ANTI-C1S ANTIBODY |
IL307501A (en) | 2021-04-19 | 2023-12-01 | Hoffmann La Roche | Modified mammalian cells |
WO2022223488A1 (en) | 2021-04-19 | 2022-10-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of splice switching oligonucleotides for exon skipping-mediated knockdown of pim2 |
WO2022223791A1 (en) | 2021-04-23 | 2022-10-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cell senescence accumulation related disease |
WO2022223651A1 (en) | 2021-04-23 | 2022-10-27 | F. Hoffmann-La Roche Ag | Prevention or mitigation of nk cell engaging agent-related adverse effects |
WO2022228705A1 (en) | 2021-04-30 | 2022-11-03 | F. Hoffmann-La Roche Ag | Dosing for combination treatment with anti-cd20/anti-cd3 bispecific antibody and anti-cd79b antibody drug conjugate |
WO2022228706A1 (en) | 2021-04-30 | 2022-11-03 | F. Hoffmann-La Roche Ag | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibody |
EP4334343A2 (en) | 2021-05-06 | 2024-03-13 | The Rockefeller University | Neutralizing anti-sars- cov-2 antibodies and methods of use thereof |
BR112023023622A2 (en) | 2021-05-12 | 2024-02-06 | Genentech Inc | METHODS TO TREAT DIFFUSE LYMPHOMA, KITS, IMMUNOCONJUGATES, POLATUZUMABE VEDOTIN AND IMMUNOCONJUGATE FOR USE |
WO2022241082A1 (en) | 2021-05-14 | 2022-11-17 | Genentech, Inc. | Agonists of trem2 |
WO2022242892A1 (en) | 2021-05-17 | 2022-11-24 | Université de Liège | Anti-cd38 single-domain antibodies in disease monitoring and treatment |
WO2022243261A1 (en) | 2021-05-19 | 2022-11-24 | F. Hoffmann-La Roche Ag | Agonistic cd40 antigen binding molecules targeting cea |
KR20240010469A (en) | 2021-05-21 | 2024-01-23 | 제넨테크, 인크. | Modified cells for production of recombinant products of interest |
EP4347657A1 (en) | 2021-05-25 | 2024-04-10 | Seagen Inc. | Methods of quantifying anti-cd40 antibodies |
EP4348257A1 (en) | 2021-05-26 | 2024-04-10 | Genentech, Inc. | Methods for modulating host cell surface interactions with human cytomegalovirus |
AR126009A1 (en) | 2021-06-02 | 2023-08-30 | Hoffmann La Roche | CD28 ANTIGEN-BINDING AGONIST MOLECULES THAT TARGET EPCAM |
EP4347653A1 (en) | 2021-06-04 | 2024-04-10 | Boehringer Ingelheim International GmbH | Anti-sirp-alpha antibodies |
CN117480184A (en) | 2021-06-04 | 2024-01-30 | 中外制药株式会社 | anti-DDR 2 antibodies and uses thereof |
WO2022258600A1 (en) | 2021-06-09 | 2022-12-15 | F. Hoffmann-La Roche Ag | Combination of a particular braf inhibitor (paradox breaker) and a pd-1 axis binding antagonist for use in the treatment of cancer |
WO2022266221A1 (en) | 2021-06-16 | 2022-12-22 | Alector Llc | Monovalent anti-mertk antibodies and methods of use thereof |
CN117642426A (en) | 2021-06-16 | 2024-03-01 | 艾莱克特有限责任公司 | Bispecific anti-MerTK and anti-PDL 1 antibodies and methods of use thereof |
TW202317625A (en) | 2021-06-17 | 2023-05-01 | 德商百靈佳殷格翰國際股份有限公司 | Novel tri-specific binding molecules |
WO2022269473A1 (en) | 2021-06-23 | 2022-12-29 | Janssen Biotech, Inc. | Materials and methods for hinge regions in functional exogenous receptors |
WO2022270611A1 (en) | 2021-06-25 | 2022-12-29 | 中外製薬株式会社 | Anti–ctla-4 antibody |
AU2022297107A1 (en) | 2021-06-25 | 2024-01-18 | Chugai Seiyaku Kabushiki Kaisha | Use of anti-ctla-4 antibody |
CA3224374A1 (en) | 2021-06-29 | 2023-01-05 | Flagship Pioneering Innovations V, Inc. | Immune cells engineered to promote thanotransmission and uses thereof |
AU2022302170A1 (en) | 2021-07-02 | 2023-12-21 | F. Hoffmann-La Roche Ag | Methods and compositions for treating cancer |
AU2022306973A1 (en) | 2021-07-09 | 2024-02-22 | Janssen Biotech, Inc. | Manufacturing methods for producing anti-il12/il23 antibody compositions |
KR20240032991A (en) | 2021-07-09 | 2024-03-12 | 얀센 바이오테크 인코포레이티드 | Manufacturing Methods for Producing Anti-TNF Antibody Compositions |
AU2022306144A1 (en) | 2021-07-09 | 2024-02-22 | Janssen Biotech, Inc. | Manufacturing methods for producing anti-tnf antibody compositions |
IL309559A (en) | 2021-07-09 | 2024-02-01 | Luxembourg Inst Of Health Lih | Dimeric protein complexes and uses thereof |
WO2023285362A1 (en) | 2021-07-12 | 2023-01-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of il-36 inhibitors for the treatment of netherton syndrome |
WO2023288241A1 (en) | 2021-07-14 | 2023-01-19 | Genentech, Inc. | Anti-c-c motif chemokine receptor 8 (ccr8) antibodies and methods of use |
CA3219606A1 (en) | 2021-07-22 | 2023-01-26 | F. Hoffmann-La Roche Ag | Heterodimeric fc domain antibodies |
WO2023004386A1 (en) | 2021-07-22 | 2023-01-26 | Genentech, Inc. | Brain targeting compositions and methods of use thereof |
AU2022317820A1 (en) | 2021-07-28 | 2023-12-14 | F. Hoffmann-La Roche Ag | Methods and compositions for treating cancer |
WO2023010080A1 (en) | 2021-07-30 | 2023-02-02 | Seagen Inc. | Treatment for cancer |
WO2023006975A2 (en) | 2021-07-30 | 2023-02-02 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Chimeric proteins and methods of immunotherapy |
CN117794953A (en) | 2021-08-03 | 2024-03-29 | 豪夫迈·罗氏有限公司 | Bispecific antibodies and methods of use |
WO2023019239A1 (en) | 2021-08-13 | 2023-02-16 | Genentech, Inc. | Dosing for anti-tryptase antibodies |
WO2023028591A1 (en) | 2021-08-27 | 2023-03-02 | Genentech, Inc. | Methods of treating tau pathologies |
TW202325727A (en) | 2021-08-30 | 2023-07-01 | 美商建南德克公司 | Anti-polyubiquitin multispecific antibodies |
TW202313695A (en) | 2021-09-15 | 2023-04-01 | 法商感應檢查療法公司 | Use of anti-btn3a antibody in manufacturing a medicament for use in treating a tumor |
WO2023052541A1 (en) | 2021-09-30 | 2023-04-06 | Imcheck Therapeutics | Combination of an anti-btn3a activating antibody and an il-2 agonist for use in therapy |
WO2023056362A1 (en) | 2021-09-30 | 2023-04-06 | Seagen Inc. | B7-h4 antibody-drug conjugates for the treatment of cancer |
WO2023056403A1 (en) | 2021-09-30 | 2023-04-06 | Genentech, Inc. | Methods for treatment of hematologic cancers using anti-tigit antibodies, anti-cd38 antibodies, and pd-1 axis binding antagonists |
WO2023060086A1 (en) | 2021-10-04 | 2023-04-13 | Takeda Vaccines, Inc. | Methods for determining norovirus-reactive antibodies |
US20230190805A1 (en) | 2021-10-06 | 2023-06-22 | Immatics Biotechnologies Gmbh | Methods of identifying metastatic lesions in a patient and treating thereof |
WO2023057601A1 (en) | 2021-10-06 | 2023-04-13 | Biotalys NV | Anti-fungal polypeptides |
WO2023062048A1 (en) | 2021-10-14 | 2023-04-20 | F. Hoffmann-La Roche Ag | Alternative pd1-il7v immunoconjugates for the treatment of cancer |
AU2022362681A1 (en) | 2021-10-14 | 2024-04-04 | F. Hoffmann-La Roche Ag | New interleukin-7 immunoconjugates |
WO2023069919A1 (en) | 2021-10-19 | 2023-04-27 | Alector Llc | Anti-cd300lb antibodies and methods of use thereof |
WO2023073084A1 (en) | 2021-10-27 | 2023-05-04 | Imcheck Therapeutics | Butyrophilin (btn) 3a activating antibodies for use in methods for treating infectious disorders |
WO2023073615A1 (en) | 2021-10-29 | 2023-05-04 | Janssen Biotech, Inc. | Methods of treating crohn's disease with anti-il23 specific antibody |
WO2023073225A1 (en) | 2021-11-01 | 2023-05-04 | F. Hoffmann-La Roche Ag | Treatment of cancer using a hla-a2/wt1 x cd3 bispecific antibody and a 4-1bb (cd137) agonist |
WO2023078900A1 (en) | 2021-11-03 | 2023-05-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating triple negative breast cancer (tnbc) |
US20230192886A1 (en) | 2021-11-08 | 2023-06-22 | Immatics Biotechnologies Gmbh | Adoptive cell therapy combination treatment and compositions thereof |
WO2023086835A1 (en) | 2021-11-09 | 2023-05-19 | Sensei Biotherapeutics, Inc. | Anti-vista antibodies and uses thereof |
WO2023086807A1 (en) | 2021-11-10 | 2023-05-19 | Genentech, Inc. | Anti-interleukin-33 antibodies and uses thereof |
US20230151087A1 (en) | 2021-11-15 | 2023-05-18 | Janssen Biotech, Inc. | Methods of Treating Crohn's Disease with Anti-IL23 Specific Antibody |
WO2023088889A1 (en) | 2021-11-16 | 2023-05-25 | Apogenix Ag | CD137 ligands |
WO2023088876A1 (en) | 2021-11-16 | 2023-05-25 | Apogenix Ag | Multi-specific immune modulators |
TW202337494A (en) | 2021-11-16 | 2023-10-01 | 美商建南德克公司 | Methods and compositions for treating systemic lupus erythematosus (sle) with mosunetuzumab |
WO2023095000A1 (en) | 2021-11-23 | 2023-06-01 | Janssen Biotech, Inc. | Method of treating ulcerative colitis with anti-il23 specific antibody |
WO2023094525A1 (en) | 2021-11-25 | 2023-06-01 | Veraxa Biotech Gmbh | Improved antibody-payload conjugates (apcs) prepared by site-specific conjugation utilizing genetic code expansion |
EP4186529A1 (en) | 2021-11-25 | 2023-05-31 | Veraxa Biotech GmbH | Improved antibody-payload conjugates (apcs) prepared by site-specific conjugation utilizing genetic code expansion |
WO2023094698A1 (en) | 2021-11-29 | 2023-06-01 | Ose Immunotherapeutics | Specific antagonist anti-sirpg antibodies |
AR127887A1 (en) | 2021-12-10 | 2024-03-06 | F Hoffmann La Roche Ag | ANTIBODIES THAT BIND CD3 AND PLAP |
WO2023110788A1 (en) | 2021-12-14 | 2023-06-22 | F. Hoffmann-La Roche Ag | Treatment of cancer using a hla-a2/mage-a4 x cd3 bispecific antibody and a 4-1bb (cd137) agonist |
WO2023110937A1 (en) | 2021-12-14 | 2023-06-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Depletion of nk cells for the treatment of adverse post-ischemic cardiac remodeling |
WO2023114543A2 (en) | 2021-12-17 | 2023-06-22 | Dana-Farber Cancer Institute, Inc. | Platform for antibody discovery |
WO2023114544A1 (en) | 2021-12-17 | 2023-06-22 | Dana-Farber Cancer Institute, Inc. | Antibodies and uses thereof |
TW202340248A (en) | 2021-12-20 | 2023-10-16 | 瑞士商赫孚孟拉羅股份公司 | Agonistic ltbr antibodies and bispecific antibodies comprising them |
WO2023118165A1 (en) | 2021-12-21 | 2023-06-29 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
EP4209508A1 (en) | 2022-01-11 | 2023-07-12 | Centre national de la recherche scientifique | Nanobodies for the deneddylating enzyme nedp1 |
WO2023141445A1 (en) | 2022-01-19 | 2023-07-27 | Genentech, Inc. | Anti-notch2 antibodies and conjugates and methods of use |
WO2023147329A1 (en) | 2022-01-26 | 2023-08-03 | Genentech, Inc. | Antibody-conjugated chemical inducers of degradation and methods thereof |
WO2023147328A1 (en) | 2022-01-26 | 2023-08-03 | Genentech, Inc. | Antibody-conjugated chemical inducers of degradation with hydolysable maleimide linkers and methods thereof |
WO2023147399A1 (en) | 2022-01-27 | 2023-08-03 | The Rockefeller University | Broadly neutralizing anti-sars-cov-2 antibodies targeting the n-terminal domain of the spike protein and methods of use thereof |
WO2023144235A1 (en) | 2022-01-27 | 2023-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for monitoring and treating warburg effect in patients with pi3k-related disorders |
WO2023144303A1 (en) | 2022-01-31 | 2023-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Cd38 as a biomarker and biotarget in t-cell lymphomas |
EP4231017A1 (en) | 2022-02-17 | 2023-08-23 | Promise Proteomics | Detection and quantification of anti-drug antibodies and anti-self antibodies |
WO2023156634A1 (en) | 2022-02-17 | 2023-08-24 | Atb Therapeutics | Recombinant immunotoxin comprising a ribosome inactivating protein |
WO2023173026A1 (en) | 2022-03-10 | 2023-09-14 | Sorrento Therapeutics, Inc. | Antibody-drug conjugates and uses thereof |
WO2023170247A1 (en) | 2022-03-11 | 2023-09-14 | Mablink Bioscience | Antibody-drug conjugates and their uses |
WO2023175171A1 (en) | 2022-03-18 | 2023-09-21 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Bk polyomavirus antibodies and uses thereof |
US20230414750A1 (en) | 2022-03-23 | 2023-12-28 | Hoffmann-La Roche Inc. | Combination treatment of an anti-cd20/anti-cd3 bispecific antibody and chemotherapy |
WO2023186756A1 (en) | 2022-03-28 | 2023-10-05 | F. Hoffmann-La Roche Ag | Interferon gamma variants and antigen binding molecules comprising these |
US20230312703A1 (en) | 2022-03-30 | 2023-10-05 | Janssen Biotech, Inc. | Method of Treating Psoriasis with IL-23 Specific Antibody |
WO2023191816A1 (en) | 2022-04-01 | 2023-10-05 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
EP4257609A1 (en) | 2022-04-08 | 2023-10-11 | iOmx Therapeutics AG | Combination therapies based on pd-1 inhibitors and sik3 inhibitors |
WO2023198648A1 (en) | 2022-04-11 | 2023-10-19 | Institut National de la Santé et de la Recherche Médicale | Methods for the diagnosis and treatment of t-cell malignancies |
TW202400243A (en) | 2022-04-12 | 2024-01-01 | 日商衛材R&D企管股份有限公司 | Eribulin-based antibody-drug conjugates and methods of use |
WO2023198727A1 (en) | 2022-04-13 | 2023-10-19 | F. Hoffmann-La Roche Ag | Pharmaceutical compositions of anti-cd20/anti-cd3 bispecific antibodies and methods of use |
WO2023201299A1 (en) | 2022-04-13 | 2023-10-19 | Genentech, Inc. | Pharmaceutical compositions of therapeutic proteins and methods of use |
WO2023198851A1 (en) | 2022-04-14 | 2023-10-19 | Institut National de la Santé et de la Recherche Médicale | Methods for controlling the tumor cell killing by light |
WO2023198874A1 (en) | 2022-04-15 | 2023-10-19 | Institut National de la Santé et de la Recherche Médicale | Methods for the diagnosis and treatment of t cell-lymphomas |
WO2023212298A1 (en) | 2022-04-29 | 2023-11-02 | Broadwing Bio Llc | Bispecific antibodies and methods of treating ocular disease |
WO2023212294A1 (en) | 2022-04-29 | 2023-11-02 | Broadwing Bio Llc | Angiopoietin-related protein 7-specific antibodies and uses thereof |
WO2023212293A1 (en) | 2022-04-29 | 2023-11-02 | Broadwing Bio Llc | Complement factor h related 4-specific antibodies and uses thereof |
WO2023215737A1 (en) | 2022-05-03 | 2023-11-09 | Genentech, Inc. | Anti-ly6e antibodies, immunoconjugates, and uses thereof |
WO2023217904A1 (en) | 2022-05-10 | 2023-11-16 | Institut National de la Santé et de la Recherche Médicale | Syncitin-1 fusion proteins and uses thereof for cargo delivery into target cells |
WO2023219613A1 (en) | 2022-05-11 | 2023-11-16 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2023218431A1 (en) | 2022-05-13 | 2023-11-16 | BioNTech SE | Rna compositions targeting hiv |
WO2023223265A1 (en) | 2022-05-18 | 2023-11-23 | Janssen Biotech, Inc. | Method for evaluating and treating psoriatic arthritis with il23 antibody |
US20230416412A1 (en) | 2022-05-31 | 2023-12-28 | Hoffmann-La Roche Inc. | Prevention or mitigation of t-cell engaging agent-related adverse effects |
WO2023240058A2 (en) | 2022-06-07 | 2023-12-14 | Genentech, Inc. | Prognostic and therapeutic methods for cancer |
WO2023237661A1 (en) | 2022-06-09 | 2023-12-14 | Institut National de la Santé et de la Recherche Médicale | Use of endothelin receptor type b agonists for the treatment of aortic valve stenosis |
EP4299124A1 (en) | 2022-06-30 | 2024-01-03 | Universite De Montpellier | Anti-mglur2 nanobodies for use as biomolecule transporter |
WO2024003310A1 (en) | 2022-06-30 | 2024-01-04 | Institut National de la Santé et de la Recherche Médicale | Methods for the diagnosis and treatment of acute lymphoblastic leukemia |
WO2024008755A1 (en) | 2022-07-04 | 2024-01-11 | Vib Vzw | Blood-cerebrospinal fluid barrier crossing antibodies |
WO2024008274A1 (en) | 2022-07-04 | 2024-01-11 | Universiteit Antwerpen | T regulatory cell modification |
WO2024008799A1 (en) | 2022-07-06 | 2024-01-11 | Institut National de la Santé et de la Recherche Médicale | Methods for the treatment of proliferative glomerulonephritis |
WO2024013234A1 (en) | 2022-07-13 | 2024-01-18 | Institut National de la Santé et de la Recherche Médicale | Methods for diagnosis, prognosis, stratification and treating of myocarditis |
WO2024015897A1 (en) | 2022-07-13 | 2024-01-18 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2024013315A1 (en) | 2022-07-15 | 2024-01-18 | Boehringer Ingelheim International Gmbh | Binding molecules for the treatment of cancer |
WO2024020432A1 (en) | 2022-07-19 | 2024-01-25 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2024018003A1 (en) | 2022-07-21 | 2024-01-25 | Institut National de la Santé et de la Recherche Médicale | Extracellular vesicles functionalized with an erv syncitin and uses thereof for cargo delivery |
WO2024018046A1 (en) | 2022-07-22 | 2024-01-25 | Institut National de la Santé et de la Recherche Médicale | Garp as a biomarker and biotarget in t-cell malignancies |
WO2024020579A1 (en) | 2022-07-22 | 2024-01-25 | Bristol-Myers Squibb Company | Antibodies binding to human pad4 and uses thereof |
WO2024018426A1 (en) | 2022-07-22 | 2024-01-25 | Janssen Biotech, Inc. | Enhanced transfer of genetic instructions to effector immune cells |
WO2024023246A1 (en) | 2022-07-28 | 2024-02-01 | Philogen S.P.A. | Antibody binding to pd1 |
WO2024026472A2 (en) | 2022-07-29 | 2024-02-01 | Alector Llc | Transferrin receptor antigen-binding domains and uses therefor |
WO2024023283A1 (en) | 2022-07-29 | 2024-02-01 | Institut National de la Santé et de la Recherche Médicale | Lrrc33 as a biomarker and biotarget in cutaneous t-cell lymphomas |
WO2024026471A1 (en) | 2022-07-29 | 2024-02-01 | Alector Llc | Cd98hc antigen-binding domains and uses therefor |
WO2024026447A1 (en) | 2022-07-29 | 2024-02-01 | Alector Llc | Anti-gpnmb antibodies and methods of use thereof |
WO2024028433A1 (en) | 2022-08-04 | 2024-02-08 | Institut National de la Santé et de la Recherche Médicale | Methods for the treatment of lymphoproliferative disorders |
WO2024033362A1 (en) | 2022-08-08 | 2024-02-15 | Atb Therapeutics | Humanized antibodies against cd79b |
WO2024033400A1 (en) | 2022-08-10 | 2024-02-15 | Institut National de la Santé et de la Recherche Médicale | Sk2 inhibitor for the treatment of pancreatic cancer |
WO2024033399A1 (en) | 2022-08-10 | 2024-02-15 | Institut National de la Santé et de la Recherche Médicale | Sigmar1 ligand for the treatment of pancreatic cancer |
WO2024040020A1 (en) | 2022-08-15 | 2024-02-22 | Absci Corporation | Quantitative affinity activity specific cell enrichment |
WO2024038112A1 (en) | 2022-08-17 | 2024-02-22 | Institut National de la Santé et de la Recherche Médicale | Improved anti-albumin nanobodies and their uses |
WO2024047110A1 (en) | 2022-08-31 | 2024-03-07 | Institut National de la Santé et de la Recherche Médicale | Method to generate more efficient car-t cells |
WO2024049949A1 (en) | 2022-09-01 | 2024-03-07 | Genentech, Inc. | Therapeutic and diagnostic methods for bladder cancer |
WO2024052503A1 (en) | 2022-09-08 | 2024-03-14 | Institut National de la Santé et de la Recherche Médicale | Antibodies having specificity to ltbp2 and uses thereof |
WO2024056668A1 (en) | 2022-09-12 | 2024-03-21 | Institut National de la Santé et de la Recherche Médicale | New anti-itgb8 antibodies and its uses thereof |
WO2024068705A1 (en) | 2022-09-29 | 2024-04-04 | F. Hoffmann-La Roche Ag | Protease-activated polypeptides |
Citations (35)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4356270A (en) * | 1977-11-08 | 1982-10-26 | Genentech, Inc. | Recombinant DNA cloning vehicle |
US4642334A (en) * | 1982-03-15 | 1987-02-10 | Dnax Research Institute Of Molecular And Cellular Biology, Inc. | Hybrid DNA prepared binding composition |
US4656134A (en) * | 1982-01-11 | 1987-04-07 | Board Of Trustees Of Leland Stanford Jr. University | Gene amplification in eukaryotic cells |
US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4704692A (en) * | 1986-09-02 | 1987-11-03 | Ladner Robert C | Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides |
US4711845A (en) * | 1984-08-31 | 1987-12-08 | Cetus Corporation | Portable temperature-sensitive control cassette |
US4714681A (en) * | 1981-07-01 | 1987-12-22 | The Board Of Reagents, The University Of Texas System Cancer Center | Quadroma cells and trioma cells and methods for the production of same |
US4800159A (en) * | 1986-02-07 | 1989-01-24 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences |
US4806471A (en) * | 1982-09-16 | 1989-02-21 | A/S Alfred Benzon | Plasmids with conditional uncontrolled replication behavior |
US4816397A (en) * | 1983-03-25 | 1989-03-28 | Celltech, Limited | Multichain polypeptides or proteins and processes for their production |
US4889818A (en) * | 1986-08-22 | 1989-12-26 | Cetus Corporation | Purified thermostable enzyme |
US4937193A (en) * | 1986-06-27 | 1990-06-26 | Delta Biotechnology Limited | Process for the genetic modification of yeast |
US4946786A (en) * | 1987-01-14 | 1990-08-07 | President And Fellows Of Harvard College | T7 DNA polymerase |
US4959317A (en) * | 1985-10-07 | 1990-09-25 | E. I. Du Pont De Nemours And Company | Site-specific recombination of DNA in eukaryotic cells |
US4965188A (en) * | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
US4978743A (en) * | 1987-09-24 | 1990-12-18 | Bayer Aktiengesellschaft | Process for the continuous, counter flow/direct flow extraction of polyamides |
US4978745A (en) * | 1987-11-23 | 1990-12-18 | Centocor, Inc. | Immunoreactive heterochain antibodies |
US4983728A (en) * | 1987-07-31 | 1991-01-08 | Ire-Celltarg S.A. | Nucleic acid probes of human papilloma virus |
US5023171A (en) * | 1989-08-10 | 1991-06-11 | Mayo Foundation For Medical Education And Research | Method for gene splicing by overlap extension using the polymerase chain reaction |
US5030565A (en) * | 1983-08-17 | 1991-07-09 | Scripps Clinic And Research Foundation | Polypeptide-induced monoclonal receptors to protein ligands |
US5091513A (en) * | 1987-05-21 | 1992-02-25 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US5126258A (en) * | 1984-09-07 | 1992-06-30 | Scripps Clinic And Research Foundation | Molecules with antibody combining sites that exhibit catalytic properties |
US5132405A (en) * | 1987-05-21 | 1992-07-21 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US5225539A (en) * | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
US5229072A (en) * | 1992-02-03 | 1993-07-20 | Liquid Carbonic Inc. | Use of interhalogen compounds as a sterilizing agent |
US5229292A (en) * | 1986-07-28 | 1993-07-20 | Stine Seed Farm, Inc. | Biological control of insects using pseudomonas strains transformed with bacillus thuringiensis insect toxingene |
US5534254A (en) * | 1992-02-06 | 1996-07-09 | Chiron Corporation | Biosynthetic binding proteins for immuno-targeting |
US5846818A (en) * | 1985-11-01 | 1998-12-08 | Xoma Corporation | Pectate lyase signal sequence |
US5885793A (en) * | 1991-12-02 | 1999-03-23 | Medical Research Council | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
US5969108A (en) * | 1990-07-10 | 1999-10-19 | Medical Research Council | Methods for producing members of specific binding pairs |
US6207804B1 (en) * | 1987-05-21 | 2001-03-27 | Curis, Inc. | Genetically engineered antibody analogues and fusion proteins thereof |
US6214553B1 (en) * | 1997-01-21 | 2001-04-10 | Massachusetts General Hospital | Libraries of protein encoding RNA-protein fusions |
US6248516B1 (en) * | 1988-11-11 | 2001-06-19 | Medical Research Council | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US6291158B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
US6291159B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
Family Cites Families (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
JPS6147500A (en) | 1984-08-15 | 1986-03-07 | Res Dev Corp Of Japan | Chimera monoclonal antibody and its preparation |
EP0173494A3 (en) | 1984-08-27 | 1987-11-25 | The Board Of Trustees Of The Leland Stanford Junior University | Chimeric receptors by dna splicing and expression |
GB8422238D0 (en) | 1984-09-03 | 1984-10-10 | Neuberger M S | Chimeric proteins |
JPS61104788A (en) | 1984-10-26 | 1986-05-23 | Teijin Ltd | Nucleic acid base sequence |
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
ES8706823A1 (en) | 1985-03-28 | 1987-06-16 | Cetus Corp | Process for amplifying, detecting, and/or cloning nucleic acid sequences. |
AU606320B2 (en) * | 1985-11-01 | 1991-02-07 | International Genetic Engineering, Inc. | Modular assembly of antibody genes, antibodies prepared thereby and use |
GB8607679D0 (en) | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
SE458555B (en) | 1986-07-21 | 1989-04-10 | Atte Heikkilae | LINFAESTANORDNING |
JPS63152984A (en) | 1986-08-18 | 1988-06-25 | Wakunaga Pharmaceut Co Ltd | Dna coding l-chain of antipyocyanic human-type antibody |
ATE87659T1 (en) * | 1986-09-02 | 1993-04-15 | Enzon Lab Inc | BINDING MOLECULES WITH SINGLE POLYPEPTIDE CHAIN. |
DE3852304T3 (en) * | 1987-03-02 | 1999-07-01 | Enzon Lab Inc | Organism as carrier for "Single Chain Antibody Domain (SCAD)". |
CA1341235C (en) | 1987-07-24 | 2001-05-22 | Randy R. Robinson | Modular assembly of antibody genes, antibodies prepared thereby and use |
CA2016841C (en) | 1989-05-16 | 1999-09-21 | William D. Huse | A method for producing polymers having a preselected activity |
EP0478627A4 (en) | 1989-05-16 | 1992-08-19 | William D. Huse | Co-expression of heteromeric receptors |
CA2016842A1 (en) | 1989-05-16 | 1990-11-16 | Richard A. Lerner | Method for tapping the immunological repertoire |
AU627591B2 (en) | 1989-06-19 | 1992-08-27 | Xoma Corporation | Chimeric mouse-human km10 antibody with specificity to a human tumor cell antigen |
EP0859841B1 (en) | 1995-08-18 | 2002-06-19 | MorphoSys AG | Protein/(poly)peptide libraries |
-
1989
- 1989-11-13 AU AU45201/89A patent/AU634186B2/en not_active Expired
- 1989-11-13 KR KR1019900701475A patent/KR0184860B1/en not_active IP Right Cessation
- 1989-11-13 EP EP89311731A patent/EP0368684B2/en not_active Expired - Lifetime
- 1989-11-13 WO PCT/GB1989/001344 patent/WO1990005144A1/en active Application Filing
- 1989-11-13 AT AT89311731T patent/ATE102631T1/en not_active IP Right Cessation
- 1989-11-13 DE DE68913658T patent/DE68913658T3/en not_active Expired - Lifetime
- 1989-11-13 JP JP1511700A patent/JP2919890B2/en not_active Expired - Lifetime
- 1989-11-13 ES ES89311731T patent/ES2052027T5/en not_active Expired - Lifetime
- 1989-11-14 CA CA002002868A patent/CA2002868C/en not_active Expired - Lifetime
-
1990
- 1990-07-09 NO NO90903059A patent/NO903059L/en unknown
- 1990-07-09 DK DK199001647A patent/DK175392B1/en not_active IP Right Cessation
- 1990-07-10 FI FI903489A patent/FI903489A0/en not_active Application Discontinuation
-
1995
- 1995-06-06 US US08/470,031 patent/US6248516B1/en not_active Expired - Lifetime
-
2000
- 2000-11-28 US US09/722,364 patent/US6545142B1/en not_active Expired - Fee Related
-
2002
- 2002-11-08 US US10/290,252 patent/US7306907B2/en not_active Expired - Fee Related
- 2002-11-08 US US10/290,233 patent/US20040110941A2/en not_active Abandoned
-
2008
- 2008-05-27 US US12/127,237 patent/US20080299618A1/en not_active Abandoned
Patent Citations (38)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4356270A (en) * | 1977-11-08 | 1982-10-26 | Genentech, Inc. | Recombinant DNA cloning vehicle |
US4714681A (en) * | 1981-07-01 | 1987-12-22 | The Board Of Reagents, The University Of Texas System Cancer Center | Quadroma cells and trioma cells and methods for the production of same |
US4656134A (en) * | 1982-01-11 | 1987-04-07 | Board Of Trustees Of Leland Stanford Jr. University | Gene amplification in eukaryotic cells |
US4642334A (en) * | 1982-03-15 | 1987-02-10 | Dnax Research Institute Of Molecular And Cellular Biology, Inc. | Hybrid DNA prepared binding composition |
US4806471A (en) * | 1982-09-16 | 1989-02-21 | A/S Alfred Benzon | Plasmids with conditional uncontrolled replication behavior |
US4816397A (en) * | 1983-03-25 | 1989-03-28 | Celltech, Limited | Multichain polypeptides or proteins and processes for their production |
US5030565A (en) * | 1983-08-17 | 1991-07-09 | Scripps Clinic And Research Foundation | Polypeptide-induced monoclonal receptors to protein ligands |
US4711845A (en) * | 1984-08-31 | 1987-12-08 | Cetus Corporation | Portable temperature-sensitive control cassette |
US5126258A (en) * | 1984-09-07 | 1992-06-30 | Scripps Clinic And Research Foundation | Molecules with antibody combining sites that exhibit catalytic properties |
US4959317A (en) * | 1985-10-07 | 1990-09-25 | E. I. Du Pont De Nemours And Company | Site-specific recombination of DNA in eukaryotic cells |
US5846818A (en) * | 1985-11-01 | 1998-12-08 | Xoma Corporation | Pectate lyase signal sequence |
US4683195B1 (en) * | 1986-01-30 | 1990-11-27 | Cetus Corp | |
US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4800159A (en) * | 1986-02-07 | 1989-01-24 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences |
US5225539A (en) * | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
US4937193A (en) * | 1986-06-27 | 1990-06-26 | Delta Biotechnology Limited | Process for the genetic modification of yeast |
US5229292A (en) * | 1986-07-28 | 1993-07-20 | Stine Seed Farm, Inc. | Biological control of insects using pseudomonas strains transformed with bacillus thuringiensis insect toxingene |
US4965188A (en) * | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
US4889818A (en) * | 1986-08-22 | 1989-12-26 | Cetus Corporation | Purified thermostable enzyme |
US4704692A (en) * | 1986-09-02 | 1987-11-03 | Ladner Robert C | Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides |
US4946786A (en) * | 1987-01-14 | 1990-08-07 | President And Fellows Of Harvard College | T7 DNA polymerase |
US6207804B1 (en) * | 1987-05-21 | 2001-03-27 | Curis, Inc. | Genetically engineered antibody analogues and fusion proteins thereof |
US5132405A (en) * | 1987-05-21 | 1992-07-21 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US5091513A (en) * | 1987-05-21 | 1992-02-25 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US4983728A (en) * | 1987-07-31 | 1991-01-08 | Ire-Celltarg S.A. | Nucleic acid probes of human papilloma virus |
US4978743A (en) * | 1987-09-24 | 1990-12-18 | Bayer Aktiengesellschaft | Process for the continuous, counter flow/direct flow extraction of polyamides |
US4978745A (en) * | 1987-11-23 | 1990-12-18 | Centocor, Inc. | Immunoreactive heterochain antibodies |
US6248516B1 (en) * | 1988-11-11 | 2001-06-19 | Medical Research Council | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US6545142B1 (en) * | 1988-11-11 | 2003-04-08 | Medical Research Council Of The United Kingdom | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US20030130496A1 (en) * | 1988-11-11 | 2003-07-10 | Medical Research Council | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US6291158B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
US6291159B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US5023171A (en) * | 1989-08-10 | 1991-06-11 | Mayo Foundation For Medical Education And Research | Method for gene splicing by overlap extension using the polymerase chain reaction |
US5969108A (en) * | 1990-07-10 | 1999-10-19 | Medical Research Council | Methods for producing members of specific binding pairs |
US5885793A (en) * | 1991-12-02 | 1999-03-23 | Medical Research Council | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
US5229072A (en) * | 1992-02-03 | 1993-07-20 | Liquid Carbonic Inc. | Use of interhalogen compounds as a sterilizing agent |
US5534254A (en) * | 1992-02-06 | 1996-07-09 | Chiron Corporation | Biosynthetic binding proteins for immuno-targeting |
US6214553B1 (en) * | 1997-01-21 | 2001-04-10 | Massachusetts General Hospital | Libraries of protein encoding RNA-protein fusions |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8940871B2 (en) | 2006-03-20 | 2015-01-27 | The Regents Of The University Of California | Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting |
US8940298B2 (en) | 2007-09-04 | 2015-01-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US9527919B2 (en) | 2007-09-04 | 2016-12-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
Also Published As
Publication number | Publication date |
---|---|
US20030130496A1 (en) | 2003-07-10 |
DE68913658T2 (en) | 1994-09-08 |
ES2052027T3 (en) | 1994-07-01 |
CA2002868A1 (en) | 1990-05-11 |
EP0368684B2 (en) | 2004-09-29 |
US6248516B1 (en) | 2001-06-19 |
KR0184860B1 (en) | 1999-04-01 |
NO903059D0 (en) | 1990-07-09 |
US7306907B2 (en) | 2007-12-11 |
AU634186B2 (en) | 1993-02-18 |
KR920700228A (en) | 1992-02-19 |
EP0368684A1 (en) | 1990-05-16 |
DK164790D0 (en) | 1990-07-09 |
JPH03502801A (en) | 1991-06-27 |
CA2002868C (en) | 2007-03-20 |
DE68913658D1 (en) | 1994-04-14 |
US6545142B1 (en) | 2003-04-08 |
ATE102631T1 (en) | 1994-03-15 |
US20030114659A1 (en) | 2003-06-19 |
ES2052027T5 (en) | 2005-04-16 |
WO1990005144A1 (en) | 1990-05-17 |
US20040110941A2 (en) | 2004-06-10 |
DK175392B1 (en) | 2004-09-20 |
JP2919890B2 (en) | 1999-07-19 |
EP0368684B1 (en) | 1994-03-09 |
NO903059L (en) | 1990-09-07 |
DK164790A (en) | 1990-09-07 |
DE68913658T3 (en) | 2005-07-21 |
AU4520189A (en) | 1990-05-28 |
FI903489A0 (en) | 1990-07-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US7306907B2 (en) | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors | |
JP3992290B2 (en) | Development of a new immunological repertoire | |
US6291158B1 (en) | Method for tapping the immunological repertoire | |
US5874060A (en) | Recombinant human anti-Lewis Y antibodies | |
US6291161B1 (en) | Method for tapping the immunological repertiore | |
US6291159B1 (en) | Method for producing polymers having a preselected activity | |
US7189841B2 (en) | Method for tapping the immunological repertoire | |
US6680192B1 (en) | Method for producing polymers having a preselected activity | |
JPH06510671A (en) | Production of chimeric antibodies - combinatorial approach | |
US20230212273A1 (en) | Chimeric antibodies for treatment of amyloid deposition diseases | |
US5976531A (en) | Composite antibodies of human subgroup IV light chain capable of binding to tag-72 | |
KR20230057351A (en) | Anti-cleavage variant CALR-CD3 bispecific antibody and pharmaceutical composition | |
US5182205A (en) | Nucleotide sequences which are selectively expressed in pre-B cells and probes therefor | |
JP2000508881A (en) | PCR amplification of rearranged genomic variable regions of immunoglobulin genes | |
EP0618969B1 (en) | Composite antibodies of human subgroup iv light chain capable of binding to tag-72 | |
Wiens et al. | Repertoire shift in the humoral response to phosphocholine-keyhole limpet hemocyanin: VH somatic mutation in germinal center B cells impairs T15 Ig function | |
WO1997031110A1 (en) | Traf family molecule, polynucleotide coding for the molecule, and antibody against the molecule | |
AU9058291A (en) | Composite antibodies of human subgroup IV light chain capable of binding to tag-72 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |
|
AS | Assignment |
Owner name: UNITED KINGDOM RESEARCH AND INNOVATION, UNITED KINGDOM Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:MEDICAL RESEARCH COUNCIL;REEL/FRAME:046469/0108 Effective date: 20180401 Owner name: UNITED KINGDOM RESEARCH AND INNOVATION, UNITED KIN Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:MEDICAL RESEARCH COUNCIL;REEL/FRAME:046469/0108 Effective date: 20180401 |