Humanized antibodies

Patent metadata

Field	Value
Publication number	US-10647756-B2
Application number	US-201615155324-A
Country	US
Kind code	B2
Filing date	May 16, 2016
Priority date	May 18, 2015
Publication date	May 12, 2020
Grant date	May 12, 2020

Abstract

Disclosed herein are humanized antibodies in which human germline residues are introduces to the complementarity determining regions (CDRs) of a non-human donor antibody. Also described herein are libraries of antibody variable domains (e.g., phage-display libraries) and methods for screening for humanized antibodies.

First claim

What is claimed is:

1. An isolated humanized monoclonal antibody or antigen-binding fragment thereof that binds to a target antigen prepared by a method comprising,
(1) obtaining the sequence of a non-human donor antibody that binds to said target antigen, and determining the donor CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 sequences of said non-human donor antibody;
(2) obtaining the sequences of a human germline VL and a human germline VH, and determining the germline framework and germline CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 sequences of said human VL and VH;
(3) aligning each of the non-human donor CDR-L1, CDR-L2, and CDR-L3 sequences with the corresponding germline CDR sequence from said human VL, and each of the non-human CDR-H1 and CDR-H2 sequences with corresponding germline CDR sequence from said human VH;
(4) identifying positions in CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 where human germline residue is the same as, or different from, the corresponding non-human donor residue; and
(5) generating the isolated humanized monoclonal antibody or antigen-binding fragment thereof by changing a non-human donor residue of the donor CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 to the corresponding human germline residue if the human germline residue at the position is different from the corresponding non-human residue, wherein:
(a) said isolated humanized monoclonal antibody or antigen-binding fragment thereof comprises (i) a VH domain comprising a human germline VH framework sequence from step (2) and a VL domain comprising a human germline VL framework sequence from step (2); and (ii) a CDR-L1, CDR-L2, CDR-L3, CDR-H1 and CDR-H2 from step (5); and (iii) a CDR-H3;
(b) for each position within said CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2, the residue is either human germline residue from said human germline VL or VH, or corresponding residue from said non-human donor antibody;
(c) said CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 each comprises at least one more human germline residue as compared to the corresponding non-human donor CDR,
(d) said CDR-L1, CDR-L2, CDR-L3, CDR-H1, and CDR-H2 each comprises at least one more non-human donor residue as compared to the corresponding human germline VH or VL CDR; and
(e) for each position within CDR-H3, the residue is any one of the 20 natural amino acid residues.

2. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence comprises a VH3, VH1, or VH5 framework sequence.

3. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence comprises a VH germline consensus framework sequence.

4. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence comprises the VH framework sequence of any one of the consensus sequences listed in Table 2 and Table 5.

5. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence comprises a VK or V?, framework sequence.

6. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence comprises a VL germline consensus framework sequence.

7. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence comprises the VL framework sequence of any one of the human germline sequences listed in Table 3, Table 4 and Table 6.

8. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence is derived from the VH framework sequence of human germline DP54.

9. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VH framework sequence is derived from the VH framework sequence of SEQ ID NO:74.

10. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence is derived from a VK framework sequence.

11. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence is derived from the VL framework sequence of human germline DPK9.

12. The antibody or antigen-binding fragment thereof of claim 1 , wherein said human germline VL framework sequence is derived from the VL framework sequence of SEQ ID NO:144.

13. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment binds said target antigen with a binding affinity (Kd) value that is equal or less than the binding affinity (Kd) value of said non-human donor antibody.

14. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment maintains highly specific binding to said target antigen as compared to binding of said non-human donor antibody to said target antigen.

Assignees

• Pfizer

Inventors

• Finlay William James Jonathan
• Tchistiakova Lioudmila
• Bennett Eric M

Classifications

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K16/40

  against enzymes · CPC title