Carbonic anhydrase IX (G250) anitbodies
The invention provides scFv antibodies and monoclonal antibodies that bind to and decrease an activity of Carbonic Anhydrase IX (G250). Also provided are methods of treating and/or preventing cancer, such as renal clear cell cancer. Also provided are methods of identifying a carbonic anhydrase IX (G250) protein. The invention additionally provides methods of modifying immune effector cells, and the immune effector cells modified thereby.
1. An isolated human monoclonal antibody that immunospecifically binds to a carbonic anhydrase IX (G250) protein, wherein said antibody
(a) has a heavy chain with a CDR1 comprising an amino acid sequence SYAMS (SEQ ID NO: 55);
(b) has a heavy chain with a CDR2 comprising an amino sequence AISXXGGXTXXADSVKG (SEQ ID NO: 58);
(c) has a heavy chain with a CDR3 comprising an amino sequence NGNYRGAFDI (SEQ ID NO: 65);
(d) has a light chain with a CDR1 comprising an amino sequence TGSSSNIGAGYDVH (SEQ ID NO: 61);
(e) has a light chain with a CDR2 comprising an amino sequence GNNNRPS (SEQ ID NO: 62); and
(f) has a light chain with a CDR3 comprising an amino sequence QSYDSSLSAWVV (SEQ ID NO: 63),
and wherein said antibody reduces carbonic anhydrase activity of said protein.
2. The antibody of claim 1 , wherein said monoclonal antibody binds to the carbonic anhydrase (CA) domain of carbonic anhydrase IX (G250).
3. An isolated scFv antibody, wherein said scFv antibody binds to a carbonic anhydrase IX (G250) protein, wherein said scFv antibody has a heavy chain comprising an amino acid sequence SEQ ID NO: 7, and wherein said scFv antibody has a light chain comprising an amino acid sequence SEQ ID NO: 28.
4. The isolated scFv antibody of claim 3 , wherein said scFv antibody binds to the carbonic anhydrase (CA) domain of a carbonic anhydrase IX (G250) protein.
5. The scFv antibody of claim 4 , wherein said binding affinity is from about 10 −6 M to about 10 −12 M.
6. An antibody complex comprising:
a) a first fully human monoclonal antibody that binds to a carbonic anhydrase IX (G250) protein, operably linked to
b) a second fully human monoclonal antibody that binds to a carbonic anhydrase IX (G250) protein, wherein said antibody complex reduces carbonic anhydrase activity, and wherein the first fully human monoclonal antibody is the antibody of claim 1 or the scFv antibody of claim 4 .
7. The antibody of claim 6 , wherein said first and second antibodies do not bind to the same epitope.
8. The monoclonal antibody of claim 1 , or a functional fragment thereof wherein said antibody contains a H310A and a H435Q mutation when numbered in accordance with the Wu and Kabat numbering system, and wherein said functional fragment binds the CA IX (G250) protein.
9. A fusion protein comprising the monoclonal antibody of claim 1 , or a functional fragment thereof, operably linked to an IL-2 polypeptide, wherein said functional fragment binds CA IX(250) protein.
10. A fusion protein comprising the monoclonal antibody of claim 1 , or a functional fragment thereof, operably linked to an IgG molecule, wherein said functional fragment binds the CA IX (G250) protein.
11. A fusion protein comprising the monoclonal antibody of claim 1 , or a functional fragment thereof, operably linked to a zeta chain of the T-cell receptor complex, wherein said functional fragment binds CA IX(250) protein.
12. The fusion protein of claim 11 , further comprising a CD28 polypeptide.
13. A minibody comprising the binding domain of the monoclonal antibody of claim 1 , or a functional fragment thereof, and a constant region of an immunoglobulin or fragment thereof, wherein said functional fragment binds the CA IX (G250) protein.
14. A composition comprising the monoclonal antibody of claim 1 and a carrier.
15. The composition of claim 14 , further comprising IL-2.
16. A kit comprising, in one or more containers, the composition of claim 14 .
17. The isolated antibody of claim 4 , wherein said antibody reduces carbonic anhydrase activity of said protein.