Patent
Antibody screening methods
| العنوان: | Antibody screening methods |
|---|---|
| Patent Number: | 9,880,160 |
| تاريخ النشر: | January 30, 2018 |
| Appl. No: | 14/005085 |
| Application Filed: | March 14, 2012 |
| مستخلص: | Provided are methods and compositions for the production of novel antibodies that bind specifically to a target antigen. These methods and compositions are particularly useful for producing antibodies having the antigen binding specificity of a reference antibody but with improved properties (e.g., binding affinity, immunogenicity, and thermodynamic stability) relative to the reference antibody. |
| Inventors: | Chen, Yan (Lexington, MA, US); Woolf, Tod M. (Sudbury, MA, US); Wagner, Richard W. (Cambridge, MA, US) |
| Assignees: | X-BODY, INC. (Waltham, MA, US) |
| Claim: | 1. A method for producing a variable heavy (VH) domain that binds a target antigen, the method comprising: (a) providing a library of chimeric, unpaired VH domains, the library comprising diversity in FR1-FR3 regions of the VH domains, and wherein each member of the library comprises a CDR3 region from the VH domain of a reference antibody that binds specifically to the target antigen; (b) contacting the library with the target antigen; and (c) selecting from the library at least one chimeric, unpaired VH domain that binds specifically to the target antigen, thereby producing a VH domain that binds specifically to the target antigen. |
| Claim: | 2. The method of claim 1 , further comprising introducing amino acid sequence diversity into the library of step (a). |
| Claim: | 3. The method of claim 1 , further comprising: (d) introducing amino acid sequence diversity into the VH domain(s) selected in step (c). |
| Claim: | 4. The method of claim 2 , wherein the amino acid sequence diversity is introduced by random mutagenesis. |
| Claim: | 5. The method of claim 1 , further comprising combining the VH domain(s) selected in step (c) with a VL domain. |
| Claim: | 6. The method of claim 1 , wherein the CDR3 region is from a rodent, lagomorph, avian, camelid, shark, or human antibody. |
| Claim: | 7. The method of claim 1 , wherein each member of the library comprises an identical CDR3 region. |
| Claim: | 8. The method of claim 1 , wherein each member of the library further comprises a human FR4 region. |
| Claim: | 9. The method of claim 1 , wherein sequences of the FR1-FR3 regions of said VH domains are human sequences. |
| Claim: | 10. The method of claim 1 , wherein each member of the library comprises FR1-FR3 regions individually encoded by a single human antibody VH gene. |
| Claim: | 11. The method of claim 1 , wherein the library is a nucleic acid display library. |
| Claim: | 12. The method of claim 1 , wherein the library is generated using the oligonucleotide sequence set forth in SEQ ID NO: 19, and at least one oligonucleotide having a sequence selected from the group consisting of SEQ ID NOs: 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, and 21. |
| Claim: | 13. The method of claim 5 , wherein the VL domain is generated using at least one oligonucleotide having a sequence selected from the group consisting of SEQ ID NOs: 70, 71, 72, 73, 74, 75, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, and 102. |
| Claim: | 14. A method for producing an antibody comprising a stable variable heavy (VH) domain/variable light (VL) domain (VH/VL) pair, the method comprising: (a) providing a library of chimeric, unpaired VH domains, the library comprising diversity in FR1-FR3 regions of the VH domains, and wherein each member of the library comprises a CDR3 region from the VH domain of a reference antibody that binds specifically to the target antigen; (b) contacting the library with the target antigen; (c) selecting from the library at least one chimeric, unpaired VH domain that binds specifically to the target antigen; and (d) contacting the unpaired VH domain selected in step (c) with a VL domain, thereby producing an antibody with a stable VH/VL pair. |
| Claim: | 15. A method for producing an antibody comprising a stable variable heavy (VH) domain/variable light (VL) domain (VH/VL) pair, the method comprising: (a) providing a library of chimeric, unpaired VH domains, the library comprising diversity in FR1-FR3 regions of the VH domains, and wherein each member of the library comprises a CDR3 region from the VH domain of a reference antibody that binds specifically to the target antigen; (b) contacting the library with the target antigen; (c) selecting from the library at least one chimeric, unpaired VH domain that binds specifically to the target antigen; (d) contacting the at least one unpaired VH domain selected in step (c) with a library of unpaired VL domains such that a library of VH/VL pairs is formed; (e) contacting the library of VH/VL pairs with the antigen; and (f) selecting from the library of VH/VL pairs at least one VH/VL pair that binds specifically to the antigen, thereby producing an antibody comprising a stable VH/VL pair. |
| Patent References Cited: | 2005/0255552 November 2005 Flynn et al. 2006/0134098 June 2006 Bebbington et al. 2010/0105569 April 2010 Hsieh 2712298 July 2009 2008/130704 October 2008 2013/085972 June 2013 |
| Other References: | International Search Report for International Application No. PCT/US2012/029086, dated Mar. 28, 2013 (5 pages). cited by applicant Written Opinion of the International Searching Authority for International Application No. PCT/US2012/029086, dated Mar. 28, 2013 (7 pages). cited by applicant Brezinschek, Hans-Peter, et al. “Pairing of variable heavy and variable κ chains in individual naive and memory B cells.” The Journal of Immunology 160.10 (1998): 4762-4767. cited by applicant Supplementary European Search Report with Written Opinion corresponding to European Patent Application No. 12757543.9, dated May 11, 2015. cited by applicant Chen et al. (2008) “Construction of a Large Phage-Displayed Human Antibody Domain Library with a Scaffold Based On a Newly Identified Highly Soluble, Stable Heavy Chain Variable Domain,” J. Mol. Biol. 382(3);779-789. cited by applicant |
| Primary Examiner: | Boesen, Christian |
| Attorney, Agent or Firm: | Lathrop Gage LLP Velema, Esq., James H. |
| رقم الانضمام: | edspgr.09880160 |
| قاعدة البيانات: | USPTO Patent Grants |
| الوصف غير متاح. |