CA2622180A1 - Method for generating f(ab')2 antibody fragments - Google Patents
Method for generating f(ab')2 antibody fragments Download PDFInfo
- Publication number
- CA2622180A1 CA2622180A1 CA002622180A CA2622180A CA2622180A1 CA 2622180 A1 CA2622180 A1 CA 2622180A1 CA 002622180 A CA002622180 A CA 002622180A CA 2622180 A CA2622180 A CA 2622180A CA 2622180 A1 CA2622180 A1 CA 2622180A1
- Authority
- CA
- Canada
- Prior art keywords
- fragment
- antibody
- acidic solution
- agent
- temperature condition
- 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.)
- Granted
Links
- 102000008394 Immunoglobulin Fragments Human genes 0.000 title 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 title 1
- 239000012634 fragment Substances 0.000 claims abstract 13
- 108090001109 Thermolysin Proteins 0.000 claims abstract 3
- 239000000203 mixture Substances 0.000 claims abstract 2
- 239000003929 acidic solution Substances 0.000 claims 7
- 239000003795 chemical substances by application Substances 0.000 claims 3
- 108060003951 Immunoglobulin Proteins 0.000 claims 2
- 239000002202 Polyethylene glycol Substances 0.000 claims 2
- 102000018358 immunoglobulin Human genes 0.000 claims 2
- 229920001223 polyethylene glycol Polymers 0.000 claims 2
- 229920000642 polymer Polymers 0.000 claims 2
- 239000003085 diluting agent Substances 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- 229940124597 therapeutic agent Drugs 0.000 claims 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/06—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from serum
- C07K16/065—Purification, fragmentation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- 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
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/54—F(ab')2
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S424/00—Drug, bio-affecting and body treating compositions
- Y10S424/809—Drug, bio-affecting and body treating compositions involving immunoglobulin or antibody fragment, e.g. fab', fv, fc, heavy chain or light chain
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/866—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof involving immunoglobulin or antibody fragment, e.g. fab', fab, fv, fc, heavy chain or light chain
Abstract
Methods for generating F(ab')2 fragments from antibodies using thermolysin as well as F(ab')2 fragments and compositions comprising F(ab')2 fragments generated by the method are described.
Claims (20)
1. A method of separating an F(ab')2 portion of an antibody from a non-F(ab')2 portion of an antibody comprising:
(a) contacting at least one antibody with thermolysin, under temperature and pH
conditions effective to cleave the F(ab')2 fragment from the non-F(ab')2 portion of the antibody; and (b) isolating the F(ab')2 fragment generated in step (a) from the non-F(ab')2 portion.
(a) contacting at least one antibody with thermolysin, under temperature and pH
conditions effective to cleave the F(ab')2 fragment from the non-F(ab')2 portion of the antibody; and (b) isolating the F(ab')2 fragment generated in step (a) from the non-F(ab')2 portion.
2. The method of claim 1, wherein the antibody is an immunoglobulin.
3. The method of claim 2, wherein the immunoglobulin is an IgG.
4. The method of claim 3, wherein the IgG is IgG1 or IgG2.
5. The method of claim 1, wherein the temperature condition of step (a) is from about 23°C to about 65°C.
6. The method of claim 1, wherein the temperature condition of step (a) is from about 42°C to about 65°C.
7. The method of claim 1, wherein the temperature condition of step (a) is from about 50°C to about 65°C.
8. The method of claim 1, wherein the temperature condition of step (a) is about 55°C.
9. The method of claim 1, wherein the pH condition of step (a) comprises an acidic solution.
10. The method of claim 9, wherein the acidic solution has a pH of about 2.0 to about 6.9.
11. The method of claim 9, wherein the acidic solution has a pH of about 3.0 to about 6Ø
12. The method of claim 9, wherein the acidic solution has a pH of about 4.0 to about 5.5.
13. The method of claim 9, wherein the acidic solution has a pH of about 4.0 to about 5Ø
14. An F(ab')2 fragment produced by the method of claim 1.
15. The F(ab')2 fragment of claim 14, further comprising a second agent.
16. The F(ab')2 fragment of claim 15, wherein the second agent is a polymer.
17. The F(ab')2 fragment of claim 16, wherein the polymer is polyethylene glycol (PEG).
18. The F(ab')2 fragment of claim 14, wherein the second agent is a therapeutic agent.
19. A composition comprising the F(ab')2 fragment of claim 14 or claim 15 and a pharmaceutically acceptable vehicle, carrier, or diluent.
20. A method of generating an F(ab')2 portion of an antibody comprising the steps of:
(a) contacting an acidic solution comprising an antibody with an effective amount of thermolysin, for a time and at a temperature effective to cleave the F(ab')2 fragment from the non-F(ab')2 portion of the antibody; and (b) isolating the F(ab')2 fragment generated in step (a) from the non-F(ab')2 components of the acidic solution.
(a) contacting an acidic solution comprising an antibody with an effective amount of thermolysin, for a time and at a temperature effective to cleave the F(ab')2 fragment from the non-F(ab')2 portion of the antibody; and (b) isolating the F(ab')2 fragment generated in step (a) from the non-F(ab')2 components of the acidic solution.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US71888305P | 2005-09-20 | 2005-09-20 | |
| US60/718,883 | 2005-09-20 | ||
| PCT/US2006/036267 WO2007035624A1 (en) | 2005-09-20 | 2006-09-18 | Method for generating f(ab')2 antibody fragments |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2622180A1 true CA2622180A1 (en) | 2007-03-29 |
| CA2622180C CA2622180C (en) | 2012-03-20 |
Family
ID=37605689
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2622180A Expired - Fee Related CA2622180C (en) | 2005-09-20 | 2006-09-18 | Method for generating f(ab')2 antibody fragments |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US7794970B2 (en) |
| EP (1) | EP1926751A1 (en) |
| AU (1) | AU2006292433B2 (en) |
| CA (1) | CA2622180C (en) |
| WO (1) | WO2007035624A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10143652B2 (en) * | 2009-09-23 | 2018-12-04 | Curirx Inc. | Methods for the preparation of liposomes |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5195125A (en) * | 1975-02-19 | 1976-08-20 | JOMYAKUCHUSHAYOMENEKIGUROBURINNO SEIZOHO | |
| DE2846412A1 (en) * | 1978-10-25 | 1980-05-08 | Behringwerke Ag | AGENT FOR TREATING ALLERGIC REACTIONS |
| JPS59186924A (en) | 1983-04-08 | 1984-10-23 | Kureha Chem Ind Co Ltd | Antitumor agent bonded with human immunoglobulin |
| US4849352A (en) * | 1984-10-09 | 1989-07-18 | Sullivan John B | Antibody purification process |
| US5576184A (en) * | 1988-09-06 | 1996-11-19 | Xoma Corporation | Production of chimeric mouse-human antibodies with specificity to human tumor antigens |
| DE69837760T3 (en) | 1997-06-13 | 2012-06-21 | Genentech, Inc. | PROTEIN PURIFICATION BY CHROMATOGRAPHY AND SUBSEQUENT FILTRATION ON LOADED LAYER |
| JP4316194B2 (en) | 2002-07-15 | 2009-08-19 | 独立行政法人科学技術振興機構 | Activated antibody enzyme and method for producing activated antibody enzyme |
-
2006
- 2006-09-18 EP EP06814847A patent/EP1926751A1/en not_active Withdrawn
- 2006-09-18 US US11/522,577 patent/US7794970B2/en active Active
- 2006-09-18 CA CA2622180A patent/CA2622180C/en not_active Expired - Fee Related
- 2006-09-18 AU AU2006292433A patent/AU2006292433B2/en not_active Ceased
- 2006-09-18 WO PCT/US2006/036267 patent/WO2007035624A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| CA2622180C (en) | 2012-03-20 |
| AU2006292433A1 (en) | 2007-03-29 |
| WO2007035624A1 (en) | 2007-03-29 |
| AU2006292433B2 (en) | 2011-05-19 |
| EP1926751A1 (en) | 2008-06-04 |
| US7794970B2 (en) | 2010-09-14 |
| US20070154471A1 (en) | 2007-07-05 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20150918 |