Methods of producing 7C10 and 16C10 CD80-specific antibodies
View Patent ↗The present invention relates to the identification of antibodies which are specific to human B7.1 antigen (CD80) and which are capable of inhibiting the binding of B7.1 to a CD28 receptor and which are not capable of inhibiting the binding of B7.1 to a CTLA-4 receptor. Two of these antibodies, 16C10 and 7C10, significantly inhibit the production of IL-2, in spite of the existence of a second activating ligand B7.2 (CD86). Blocking of the primary activation signal between CD28 and B7.1 (CD80) with these antibodies while allowing the unimpaired or coincident interaction of CTLA-4 and B7.1 and/or B7.2 represents a combined antagonistic effect on positive co-stimulation with an agonistic effect on negative signalling. These antibodies may be used as specific immunosuppressants, e.g., for the treatment of autoimmune diseases and to prevent organ transplant rejection.
1. A method of producing a monoclonal anti-CD80 antibody or CD80 binding fragment thereof comprising:
(a) expressing a nucleic acid molecule in a transfectoma, wherein the nucleic acid encodes an anti-CD80 antibody or binding fragment thereof that binds specifically to human CD80 antigen and comprises variable regions of an antibody produced by a hybridoma assigned ATCC Accession No. HB-12117, or variable regions of an antibody, produced by a hybridoma assigned ATCC Accession No. HB-12119, and
(b) recovering the anti-CD80 antibody or CD80-binding fragment.
2. The method of claim 1 , wherein the nucleic acid encodes an antibody with human light chain and heavy chain constant regions.
3. The method of claim 2 , wherein the nucleic acid encodes an antibody comprising a human heavy chain constant region selected from the group consisting of a human gamma 1 constant region, a human gamma 4 constant region, and a human gamma 4 PE constant region.
4. The method of claim 1 , wherein the nucleic acid encodes an antibody comprising variable regions of a 16C10 antibody, produced by a hybridoma assigned ATCC Accession No. HB-12119 and a human gamma 1 constant region.
5. The method of claim 1 , wherein the nucleic acid encodes an antibody fragment selected from the group consisting of Fab, F(ab′) 2 and Fv.
6. A method of producing a monoclonal anti-CD80 antibody or CD80 binding fragment thereof comprising:
(a) expressing a nucleic acid molecule in a transfectoma, wherein the nucleic acid molecule encodes a monoclonal antibody which specifically binds to human CD80 antigen, wherein the antibody comprises the light chain and heavy chain variable regions contained in SEQ ID NO: 2 and SEQ ID NO: 4, respectively, and human light chain and heavy chain constant regions; and
(b) recovering the anti-CD80 antibody or CD80-binding fragment.
7. The method of claim 6 , wherein then nucleic acid encoding the light chain and heavy chain variable regions comprises a nucleotide sequence contained in SEQ ID NO: 1 and SEQ ID NO:3, respectively.
8. The method of claim 6 , wherein the nucleic acid encodes the light chain and heavy chain set forth as SEQ ID NO: 2 and SEQ ID NO: 4, respectively.
9. The method of claim 8 , wherein the nucleic acid comprises the nucleotide sequences set forth as SEQ ID NO: 1 and SEQ ID NO: 3, respectively.
10. The method of claim 6 , wherein the human heavy chain constant region is a human gamma 1 constant region, human gamma 4 constant region, or human gamma 4 PE constant region.
11. A method of producing a monoclonal anti-CD80 antibody or CD80 binding fragment thereof comprising:
(a) expressing a nucleic acid molecule in a transfectoma, wherein the nucleic acid molecule encodes a primatized monoclonal antibody which specifically binds to human CD80 antigen, wherein the antibody comprises the light chain and heavy chain variable regions contained in SEQ ID NO: 10 and SEQ ID NO: 12, respectively, and human light chain and heavy chain constant regions; and
(b) recovering the anti-CD80 antibody or CD80-binding fragment.
12. The method of claim 11 , wherein then nucleic acid encoding the light chain and heavy chain variable regions comprises a nucleotide sequence contained in SEQ ID NO: 9 and SEQ ID NO:11, respectively.
13. The method of claim 11 , wherein the nucleic acid encodes the light chain and heavy chain set forth as SEQ ID NO: 10 and SEQ ID NO: 12, respectively.
14. The method of claim 13 , wherein the nucleic acid comprises the nucleotide sequences set forth as SEQ ID NO: 9 and SEQ ID NO: 11, respectively.
15. The method of claim 11 , wherein the human heavy chain constant region is a human gamma 1 constant region, human gamma 4 constant region, or human gamma 4 PE constant region.
16. A method of producing a monoclonal anti-CD80 antibody or CD80 binding fragment thereof comprising:
(a) expressing a nucleic acid molecule in a transfectoma, wherein the nucleic acid molecule encodes a monoclonal antibody, wherein the antibody specifically binds to human CD80 antigen and comprises a light chain and heavy chain variable region of an antibody produced by a hybridoma assigned ATCC Accession No. HB-12117 and human heavy chain and light chain constant regions; and
(b) recovering the anti-CD80 antibody or CD80-binding fragment.
17. The method of claim 16 , wherein the nucleic acid encodes the antibody produced by a hybridoma assigned ATCC Accession No. HB-12117.
18. The method of claim 17 , wherein the nucleic acid comprises a nucleotide sequence encoding an anti-CD80 antibody and expressed by a hybridoma assigned ATCC Accession No. HB-12117.
19. The method of claim 16 , wherein the human heavy chain constant region is a human gamma 1 constant region, a human gamma 4 constant region, or a human gamma 4 PE constant region.
20. A method of producing a monoclonal anti-CD80 antibody or CD80 binding fragment thereof comprising:
(a) expressing a nucleic acid molecule in a transfectoma, wherein the nucleic acid molecule encodes a monoclonal antibody, wherein the antibody specifically binds to human CD80 antigen and comprises a light and heavy chain variable region of an antibody produced by a hybridoma assigned ATCC Accession No. HB-12119 and human heavy chain and light chain constant regions; and
(b) recovering the anti-CD80 antibody or CD80-binding fragment.
21. The method of claim 20 , wherein the nucleic acid encodes the antibody produced by a hybridoma assigned ATCC Accession No. HB-12119.
22. The method of claim 21 , wherein the nucleic acid comprises a nucleotide sequence encoding an anti-CD80 antibody and expressed by a hybridoma assigned ATCC Accession No. HB-12117.
23. The method of claim 20 , wherein the human heavy chain constant region is a human gamma 1 constant region, a human gamma 4 constant region, or a human gamma 4 PE constant region.