WO1991002981A1 - Assay method - Google Patents
Assay method Download PDFInfo
- Publication number
- WO1991002981A1 WO1991002981A1 PCT/GB1990/001319 GB9001319W WO9102981A1 WO 1991002981 A1 WO1991002981 A1 WO 1991002981A1 GB 9001319 W GB9001319 W GB 9001319W WO 9102981 A1 WO9102981 A1 WO 9102981A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- analyte
- ligand
- receptor pair
- metal surface
- antibody
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
-
- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6825—Nucleic acid detection involving sensors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
- G01N33/553—Metal or metal coated
Definitions
- This invention concerns a method of assaying for a macromolecular analyte by use of the technique of surface plasmon resonance spectrometry (SPRS) .
- SPRS surface plasmon resonance spectrometry
- the intensity of monochromatic plain-polarised light (conveniently obtained from a laser) reflected from the interface between an optically transparent material, e.g. glass, and a thin metal layer depends on the refractive index of material on the downstream side of the metal. Accordingly, by measuring changes in intensity of the reflected light an indication can be obtained of changes in refractive index of material at a particular point on the down-stream surface of the metal.
- the intensity of reflected light also varies with the angle of incidence, and reflectivity drops sharply to a minimum at a particular angle which is characteristic of the equipment.
- WO 89/08260 describes a method of analysing for an analyte in a ⁇ ...nple by bringing the sample into contact with a metal surface, on which an antibodv has previously been reversibly bound to immobilised analyte or analogue, and monitoring displacement of antibody as indicative of the presence or the concentration of the analyte in the sample. That specification is mainly concerned with hapten analytes and describes only hapten-antibody and antigen- antibody binding pairs.
- EPA 276142 describes competition assays involving an analyte and two other reagents, in which SPRS is used to monitor the formation of a complex on a solid surface.
- the assay involves the use of at least one other liquid reagent in addition to the sample.
- the signal resulting from complex formation on the surface may be obscured by noise resulting from non-specific binding of other macromolecules to the surface.
- This invention provides a method of assaying for an analyte, preferably a macromolecular analyte, which is a member of a ligand-receptor pair other than a hapten-antibody or an antigen-antibody pair, by the use of a metal surface adapted for surface plasmon resonance spectrometry which metal surface carries the analyte or an analogue thereof immobilised thereon with the other member of the ligand-receptor pair reversibly bound thereto, which method comprises bringing a fluid containing the analyte into contact with the metal surface and observing by surface plasmon resonance spectrometry displacement of the other member of the ligand-receptor pair from the surface.
- the analyte is a member of a ligand- receptor pair. Many example ⁇ of such pairs are known and include the following:
- protein is used to 0 include peptides.
- the method involves the use of a metal surface adapted for surface plasmon resonance spectrometry.
- the metal may comprise silver or gold, conveniently in the form of a layer e.g. deposited by evaporation on a 5 carrier such as a glass slide.
- the metal surface carries the analyte or an analogue thereof immobilised thereon.
- An analogue of the analyte is a substance which competes with the analyte for binding to the other member of the ligand-receptor pair. Often the 0 analogue will be arranged to be as near as possible or even completely identical to the analyte.
- the use in assays of analyte analogues is well known.
- Binding of the analyte or analogue to the metal surface without loss of binding power is effected by methods which are well known. Particularly when the analyte or analogue is of low molecular weight, the use of a spacer molecule may be required. To prevent non ⁇ specific binding at a later stage in the assay, any surplus area of the metal surface may be coated e.g. ° with an inert protein.
- this invention provides an assay device comprising a metal surface adapted for 5 surface plasmon resonance spectrometry, which surface' carries immobilised thereon a member of a ligand- receptor pair, other than a hapten-antibody or an antigen-antibody pair, with the other member of the ligand-receptor pair reversibly bound thereto.
- That other member of the ligand-receptor pair may have been modified to increase the signal generated to increase the SPR signal generated by its removal from the metal surface. Such modification may involve addition of a molecule or group of low refractive index, or more preferably high refractive index such as polystyrene, titanium dioxide or gold colloid.
- the method of the invention is very simple.
- a fluid containing the analyte is brought into contact with the pre-coated metal surface.
- Analyte in the fluid sample competes with immobilised analyte for binding to the other member of the ligand-receptor pair.
- Displacement of the other member of the ligand-receptor pair into solution, as a complex with analyte in the sample, is monitored by surface plasmon resonance spectrometry.
- the method has two particular advantages: (a) The only fluid reagent involved is the sample containing the analyte. When this is brought into contact with the coated metal surface, the presence or the concentration of the analyte in the sample can be assayed within minutes or even seconds. the SPRS signal, generated by removal of a reagent from the metal surface, is not significantly contaminated by noise due to non-specific binding of material to the surface.
- oligonucleotide A (97mer) was hybridised to a complementary oligonucleotide B (17mer). Oligonucleotide B was then covalently linked to the silver surface of a slide. The slide was then blocked with hybridisation buffer. A further oligonucleotide C (50mer) which had the same sequence as part of oligonucleotide B and therefore complementary to A, was then added and displacement from the silver slide of the hybridised oligonucleotide A was measured. Results using 32P end-labelled oligonucleotide A showed that as the amount of oligonucleotide C was increased more oligonucleotide A was released. Such changes will also be measurable by SPR.
- oli ⁇ onucleotides A sixteen mer probe and a complementary ninety-seven mer target were prepared using phosphoramidite chemistry on the Applied Biosystems Model 380D DNA synthesizer.
- the DNA probe was modified in order to permit efficient and stable binding to the ver surface. This was prepared by attaching a terminal primary amine at the 5 ' -end of the molecule.
- the amino-oligonucleotide was mixed with an equal volume of 0.1M sodium hydrogen carbonate solution pH 8.5 and to this mixture a solution of SPDP (0.25 mg for each 1.0 OD of oligo used) in DMF was added.
- reaction was left to equilibriate for 90 minutes at room temperature and then eluted through a Sephadex G25 PD10 column.
- Fractions containing the required product were pooled together and purified by preparative HPLC. After purification the appropriate fractions were dried down by vacuum centrifugation to remove HPLC solvents and then reconstituted in a known volume of water.
- DNA hybrid consisting of the modified 16 mer and a complementary 97 mer was prepared by mixing 16 mer and 97 mer in a 1.5:1 molar ratio ensuring that the
- the immobilisable probe 16 mer, the immobilisable probe, was in excess.
- the preannealing was carried out at 65°C in 2x SSPE (300mM NaC1, 20mM NaH.PO.-H ⁇ , 2mM ethylenediaminetetra- acetic acid) and the mixture was allowed to cool down to room temperature for 2hrs.
- Results demonstrate displacement of the 97 mer sequence away from the silver surface by the complementary 50 base sequence. This does not occur in the absence of this oligonucleotide nor in the presence of the non-complementary sequence which does not hybridise to the 97 mer.
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB8919411.2 | 1989-08-25 | ||
GB898919411A GB8919411D0 (en) | 1989-08-25 | 1989-08-25 | Assay method |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1991002981A1 true WO1991002981A1 (en) | 1991-03-07 |
Family
ID=10662144
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/GB1990/001319 WO1991002981A1 (en) | 1989-08-25 | 1990-08-24 | Assay method |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP0489074A1 (en) |
JP (1) | JPH05500109A (en) |
CA (1) | CA2063707A1 (en) |
GB (1) | GB8919411D0 (en) |
WO (1) | WO1991002981A1 (en) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993006241A1 (en) * | 1991-09-16 | 1993-04-01 | The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland | Gene probe biosensor method |
WO1993025909A1 (en) * | 1992-06-11 | 1993-12-23 | Pharmacia Biosensor Ab | Improvements in or relating to analyte detection |
DE19927051A1 (en) * | 1999-06-14 | 2000-12-21 | November Ag Molekulare Medizin | Method and device for identifying a nucleotide sequence |
WO2001031057A2 (en) * | 1999-10-22 | 2001-05-03 | Nanogen Recognomics Gmbh | Double-strand nucleic acid probes and the use thereof |
NL1014816C2 (en) * | 2000-03-31 | 2001-10-02 | Tno | Reversibly binding receptor to sensor surface, useful e.g. for identifying or isolating specific ligands, by attaching to coupling agent that binds to antibody on the surface |
WO2001079848A1 (en) * | 2000-03-31 | 2001-10-25 | Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno | Method for determination of binding with natural receptors |
WO2008105918A2 (en) * | 2006-08-18 | 2008-09-04 | 3M Innovative Properties Company | Methods of detection using acousto-mechanical detection systems |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11332595A (en) * | 1997-07-09 | 1999-12-07 | Masao Karube | Detection of dna with probe pna |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4626513A (en) * | 1983-11-10 | 1986-12-02 | Massachusetts General Hospital | Method and apparatus for ligand detection |
WO1987003093A1 (en) * | 1985-11-18 | 1987-05-21 | Radiometer A/S | Sensor for determining the concentration of a biochemical species |
EP0245206A1 (en) * | 1986-05-05 | 1987-11-11 | IntraCel Corporation | Analytical method for detecting and measuring specifically sequenced nucleic acid |
EP0276142A1 (en) * | 1987-01-21 | 1988-07-27 | ARS Holding 89 N.V. | Assay technique |
WO1989008260A1 (en) * | 1988-02-27 | 1989-09-08 | Amersham International Plc | Immobilisation of haptens and measurement |
WO1989009408A1 (en) * | 1988-03-29 | 1989-10-05 | Ares-Serono Research & Development Limited Partner | Method of assay |
-
1989
- 1989-08-25 GB GB898919411A patent/GB8919411D0/en active Pending
-
1990
- 1990-08-24 WO PCT/GB1990/001319 patent/WO1991002981A1/en not_active Application Discontinuation
- 1990-08-24 CA CA002063707A patent/CA2063707A1/en not_active Abandoned
- 1990-08-24 JP JP2512188A patent/JPH05500109A/en active Pending
- 1990-08-24 EP EP90912817A patent/EP0489074A1/en not_active Withdrawn
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4626513A (en) * | 1983-11-10 | 1986-12-02 | Massachusetts General Hospital | Method and apparatus for ligand detection |
WO1987003093A1 (en) * | 1985-11-18 | 1987-05-21 | Radiometer A/S | Sensor for determining the concentration of a biochemical species |
EP0245206A1 (en) * | 1986-05-05 | 1987-11-11 | IntraCel Corporation | Analytical method for detecting and measuring specifically sequenced nucleic acid |
EP0276142A1 (en) * | 1987-01-21 | 1988-07-27 | ARS Holding 89 N.V. | Assay technique |
WO1989008260A1 (en) * | 1988-02-27 | 1989-09-08 | Amersham International Plc | Immobilisation of haptens and measurement |
WO1989009408A1 (en) * | 1988-03-29 | 1989-10-05 | Ares-Serono Research & Development Limited Partner | Method of assay |
Cited By (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993006241A1 (en) * | 1991-09-16 | 1993-04-01 | The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland | Gene probe biosensor method |
GB2274710A (en) * | 1991-09-16 | 1994-08-03 | Secr Defence | Gene probe biosensor method |
GB2274710B (en) * | 1991-09-16 | 1995-09-06 | Secr Defence | Detection of nucleic acid sequences in a sample using an oligonucleotide immobilised on a waveguide |
US5750337A (en) * | 1991-09-16 | 1998-05-12 | The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland | Methods for detecting nucleic acid sequences using evanescent wave detection |
WO1993025909A1 (en) * | 1992-06-11 | 1993-12-23 | Pharmacia Biosensor Ab | Improvements in or relating to analyte detection |
US5965456A (en) * | 1992-06-11 | 1999-10-12 | Biacore Ab | Analyte detection |
DE19927051A1 (en) * | 1999-06-14 | 2000-12-21 | November Ag Molekulare Medizin | Method and device for identifying a nucleotide sequence |
DE19927051C2 (en) * | 1999-06-14 | 2002-11-07 | November Ag Molekulare Medizin | Method and device for identifying a nucleotide sequence |
US7067253B1 (en) | 1999-06-14 | 2006-06-27 | november Aktiengesellschaft Gesellschaft für Molekulare Medizin | Method and device for identifying a polymer |
WO2001031057A2 (en) * | 1999-10-22 | 2001-05-03 | Nanogen Recognomics Gmbh | Double-strand nucleic acid probes and the use thereof |
WO2001031057A3 (en) * | 1999-10-22 | 2001-12-27 | Aventis Res & Tech Gmbh & Co | Double-strand nucleic acid probes and the use thereof |
NL1014816C2 (en) * | 2000-03-31 | 2001-10-02 | Tno | Reversibly binding receptor to sensor surface, useful e.g. for identifying or isolating specific ligands, by attaching to coupling agent that binds to antibody on the surface |
WO2001079848A1 (en) * | 2000-03-31 | 2001-10-25 | Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno | Method for determination of binding with natural receptors |
WO2008105918A2 (en) * | 2006-08-18 | 2008-09-04 | 3M Innovative Properties Company | Methods of detection using acousto-mechanical detection systems |
WO2008105918A3 (en) * | 2006-08-18 | 2008-11-13 | 3M Innovative Properties Co | Methods of detection using acousto-mechanical detection systems |
Also Published As
Publication number | Publication date |
---|---|
GB8919411D0 (en) | 1989-10-11 |
EP0489074A1 (en) | 1992-06-10 |
JPH05500109A (en) | 1993-01-14 |
CA2063707A1 (en) | 1991-02-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6197515B1 (en) | Molecular recognition at surfaces derivatized with self-assembled monolayers | |
AU742122B2 (en) | Capacity affinity sensor | |
AU711956B2 (en) | Biosensor device and method | |
US7087148B1 (en) | Binding acceleration techniques for the detection of analytes | |
US5753518A (en) | Method of determining affinity and kinetic properties | |
US6720177B2 (en) | Porous semiconductor-based optical interferometric sensor | |
US5658732A (en) | Assay method for biological target complexes on the surface of a biosensor | |
EP1075656B1 (en) | Detection of a target in a sample | |
Henke et al. | Covalent immobilization of single-stranded DNA onto optical fibers using various linkers | |
AU2003247788A1 (en) | Nanoparticle polyanion conjugates and methods of use thereof in detecting analytes | |
US6472148B1 (en) | Molecular recognition at surfaces derivatized with self-assembled monolayers | |
JP3127301B2 (en) | Analysis technology | |
Lillis et al. | Dual polarisation interferometry characterisation of DNA immobilisation and hybridisation detection on a silanised support | |
WO1991002981A1 (en) | Assay method | |
Storri et al. | A piezoelectric biosensor for DNA hybridisation detection | |
EP1739427A1 (en) | Measurement method using biosensor | |
EP1462528A1 (en) | Nucleic acid immobilization and use as a biosensor | |
EP1606620B1 (en) | Immobilization method and kit therefor | |
JP4193039B2 (en) | Double-stranded oligonucleotide array | |
Neumann et al. | Mismatch discrimination in oligonucleotide hybridization reactions using single strand binding protein. A surface plasmon fluorescence study | |
Vandenberg et al. | Immobilization of proteins for biosensor development | |
JP3998977B2 (en) | Detection method for biochemical samples |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): CA JP US |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB IT LU NL SE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1990912817 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2063707 Country of ref document: CA |
|
WWP | Wipo information: published in national office |
Ref document number: 1990912817 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1990912817 Country of ref document: EP |