Humanized rabbit antibodies

• US 20040086979A1
• Filed: 08/07/2003
• Published: 05/06/2004
• Est. Priority Date: 08/15/2002
• Status: Abandoned Application

• Current Assignee: Epitomics Incorporated (Abcam Plc)
• Original Assignee: Epitomics Incorporated (Abcam Plc)
• Inventors: Yu, Guoliang, Pytela, Robert, Couto, Fernando Jose Rebelo Do, Zhang, Dongxiao
• Application Number:
  US10/638,210
• Publication Number:
  US 20040086979A1
• Time in Patent Office: Days
• US Class Current: 435/70.21
• CPC Class Codes: C07K 16/2839 against the integrin superfamily
• CPC Class Codes: C07K 16/467 Igs with modifications in the FR-residues only
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered

Abstract

The invention provides methods for producing a modified nucleic acid that encodes a modified a rabbit antibody so that the modified rabbit antibody is less immunogenic in a non-rabbit host than an unmodified parent rabbit antibody. The invention further provides modified nucleic acids made by these methods, as well as vectors and host cells comprising the nucleic acids, and methods for producing the encoded modified antibodies. Also provided are modified rabbit antibodies encoded by subject nucleic acids, and compositions containing the same. The invention further provides kits for carrying out the subject methods.

Claims

1. A method for resurfacing a rabbit antibody, said method comprising:

(a) identifying a surface-exposed amino acid of a framework region of a parent rabbit antibody that differs from an amino acid at a corresponding position of a non-rabbit antibody by comparing the amino acid sequence of said framework region of said parent rabbit antibody to the amino acid sequence of said framework region of said non-rabbit antibody; and

(b) substituting said identified amino acid with an amino acid at said corresponding position of a non-rabbit antibody,

to resurface said rabbit antibody.

2. The method of claim 1, further comprising identifying amino acids of said framework region of a parent antibody that are proximal to a CDR and substituting only those amino acids that are not proximal to said CDR.

3. The method of claim 1, further comprising identifying amino acids in a D-E loop region of said parent antibody and substituting only those amino acids that are not in said D-E loop.

4. The method of claim 1, wherein said identifying step involves molecular modeling of said parent rabbit antibody or said non-rabbit antibody to identify said surface-exposed amino acids.

5. The method according to claim 1, wherein said identifying step (a) comprises identifying a plurality of amino acids and step (b) comprises substituting said plurality of amino acids.

6. The method according to claim 5, wherein said plurality of amino acids is at least two discontiguous amino acid.

7. The method according to claim 1, wherein said method is a method of humanizing a rabbit monoclonal antibody.

8. The method according to claim 1, wherein a modified rabbit antibody comprising a framework region having said substituted amino acid is less immunogenic in a non-rabbit host than said rabbit parent antibody.

9. The method according to claim 1, wherein said identifying step (a) comprises identifying amino acids that may be inserted into or deleted from said framework region of said parent rabbit antibody and said substituting step (b) comprises inserting into or deleting said amino acids from said nucleic acid sequence.

10. The method according to claim 1, wherein said rabbit antibody from a rabbit of known V_H allotype.

11. The nucleic acid according to claim 10, wherein said rabbit antibody is from a rabbit that is homozygous for a VH allotype.

12. The nucleic acid according to claim 19, wherein said rabbit is homozygous for an allotype chosen from V_H1-a1, V_H1-a2 and V_H1-a3.

13. The method according to claim 1, wherein said resurfaced antibody has a binding affinity of 10⁸ M⁻

1 or greater for a specific antigen.

14. A monoclonal antibody that has been resurfaced by the method set forth in claim 1.

15. A nucleic acid encoding the monoclonal antibody of claim 14.

16. A vector comprising the nucleic acid of claim 15.

17. A host cell comprising the vector according to claim 16.

18. A method of producing a modified rabbit antibody that is less immunogenic in a non-rabbit host as compared to its parent rabbit antibody, said method comprising:

incubating the host cell of claim 17 under conditions sufficient to produce said antibody; and

harvesting said antibody.

19. A computer-readable medium encoding instructions to direct a machine to perform the method of claim 1.

20. A kit for use in a computer, said kit comprising:

(a) a computer-readable medium according to claim 19;

and

(b) instructions for operating said computer according to said programming.