DNA encoding antibodies with altered effector functions

• US 5,648,260 A
• Filed: 06/07/1995
• Issued: 07/15/1997
• Est. Priority Date: 03/18/1987
• Status: Expired due to Term

• Current Assignee: SB2 Incorporated
• Original Assignee: Scotgen Biopharmaceuticals, Inc.
• Inventors: Duncan, Alexander Robert, Burton, Dennis Raymond, Winter, Gregory Paul
• Primary Examiner(s): Feisee, Lila
• Assistant Examiner(s): Reeves, Julie E.
• Application Number:
  US08/478,825
• Time in Patent Office: 769 Days
• Field of Search: 435/252.3, 435/320.1, 435/69.6, 536/23.53
• US Class Current: 435/252.3
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002

Abstract

The DNA encoding an antibody with an altered function, e.g. altered affinity for an effector ligand such as Fc receptor (FcR) on a cell or the Cl component of complement is produced by replacing the nucleic acid encoding at least one amino acid residue in the constant portion of the antibody with nucleic acid encoding a different residue.

Claims

1. Isolated DNA encoding at least part of a constant region of an immunoglobulin heavy chain in which at least one amino acid selected from the group consisting of amino acid residues 234, 235, 236, 237, 297, 318, 320 and 322 is substituted with an amino acid which is different from that present in an unmodified antibody thereby causing an alteration in an effector function while retaining binding to antigen as compared with the unmodified antibody.

2. The isolated DNA of claim 1, wherein at least one amino acid selected from the group consisting of amino acid residues 234, 235, 236 and 237 of said immunoglobulin heavy chain is substituted with an amino acid which is different from that in an unmodified antibody, thereby causing a reduction in binding affinity for a Fc receptor as compared to the unmodified antibody.

3. The isolated DNA of claim 2, wherein at least one of residues 234, 236 and 237 of said immunoglobulin heavy chain is substituted with alanine.

4. The isolated DNA of claim 2, wherein residue 235 of said immunoglobulin heavy chain is substituted with glutamine.

5. The isolated DNA of claim 1, wherein amino acid residue 297 of said immunoglobulin heavy chain is substituted with an amino acid which is different from that present in an unmodified antibody, thereby causing a decrease in lytic properties as compared with an unmodified antibody.

6. The isolated DNA of claim 5, wherein residue 297 of said immunoglobulin heavy chain is substituted with alanine.

7. The isolated DNA of claim 1, wherein at least one amino acid selected from the group consisting of amino acid residues 318, 320 and 322 of said immunoglobulin heavy chain is substituted with an amino acid which is different from that in an unmodified antibody, thereby causing a reduction in binding affinity for Clq as compared with unmodified antibody.

8. The isolated DNA of claim 7, wherein at least one of residues 318, 320 and 322 of said immunoglobulin heavy chain is substituted with alanine, thereby reducing Clq binding affinity.

9. The isolated DNA of claim 7, wherein residue 318 of said immunoglobulin heavy chain is substituted with valine.

10. The isolated DNA of claim 7, wherein residue 322 of said immunoglobulin heavy chain is substituted with glutamine.

11. A replicable expression vector comprising a suitable promoter operably linked to a DNA which encodes at least part of a constant region of an immunoglobulin heavy chain in which at least one amino acid selected from the group consisting of amino acid residues 234, 235, 236, 237, 297, 318, 320 and 322 is substituted with an amino acid which is different from that present in an unmodified antibody thereby causing an alteration in an effector function while retaining binding to antigen as compared with the unmodified antibody.

12. The replicable expression vector of claim 11, wherein at least one amino acid selected from the group consisting of amino acid residues 234, 235, 236 and 237 of said immunoglobulin heavy chain is substituted with an amino acid which is different from that in an unmodified antibody, thereby causing a reduction in binding affinity for a Fc receptor as compared to the unmodified antibody.

13. The replicable expression vector of claim 12, wherein at least one of residues 234, 236 and 237 of said immunoglobulin heavy chain is substituted with alanine.

14. The replicable expression vector of claim 12, wherein residue 235 of said immunoglobulin heavy chain is substituted with glutamine.

15. The replicable expression vector of claim 11, wherein amino acid residue 297 of said immunoglobulin heavy chain is substituted with an amino acid which is different from that present in an unmodified antibody, thereby causing a reduction in lyric properties as compared with an unmodified antibody.

16. The replicable expression vector of claim 15, wherein residue 297 of said immunoglobulin heavy chain is substituted with alanine.

17. The replicable expression vector of claim 11, wherein at least one amino acid selected from the group consisting of amino acid residues 318, 320 and 322 of said immunoglobulin heavy chain is substituted with an amino acid which is different from that in an unmodified antibody, thereby causing a reduction in binding affinity for Clq as compared with unmodified antibody.

18. The replicable expression vector of claim 17, wherein at least one of residues 318, 320 and 322 of said immunoglobulin heavy chain is substituted with alanine, thereby reducing Clq binding affinity.

19. The replicable expression vector of claim 17, wherein residue 318 of said immunoglobulin heavy chain is substituted with valine.

20. The replicable expression vector of claim 17, wherein residue 322 of said immunoglobulin heavy chain is substituted with glutamine.

21. A transformed host cell comprising the replicable expression vector of claim 11.

22. A transformed host cell comprising the replicable expression vector of claim 12.

23. A transformed host cell comprising the replicable expression vector of claim 13.

24. A transformed host cell comprising the replicable expression vector of claim 14.

25. A transformed host cell comprising the replicable expression vector of claim 15.

26. A transformed host cell comprising the replicable expression vector of claim 16.

27. A transformed host cell comprising the replicable expression vector of claim 17.

28. A transformed host cell comprising the replicable expression vector of claim 18.

29. A transformed host cell comprising the replicable expression vector of claim 19.

30. A transformed host cell comprising the replicable expression vector of claim 20.