Anti-polyubiquitin antibodies and methods of use
The invention provides anti-polyubiquitin antibodies and methods of using the same.
1. An isolated monoclonal antibody that binds K11-linked polyubiquitin, wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 2 and 57 to 60, an HVR-L2 sequence selected from SEQ ID NOs: 3 and 61, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence selected from SEQ ID NOs: 6 to 11, an HVR-H2 sequence selected from SEQ ID NOs: 12 to 17 and 67, and an HVR-H3 sequence selected from SEQ ID NOs: 18 to 23, 68 and 69.
2. The antibody of claim 1 , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 2 and 57 to 60, an HVR-L2 sequence selected from SEQ ID NOs: 3 and 61, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence of SEQ ID NO: 11, an HVR-H2 sequence selected from SEQ ID NOs: 17 and 67, and an HVR-H3 sequence selected from SEQ ID NOs: 23, 68 and 69.
3. The antibody of claim 1 , wherein the antibody comprises an HVR-L1 sequence selected from SEQ ID NOs: 58 and 59, an HVR-L2 sequence of SEQ ID NO: 3, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence of SEQ ID NO: 11, an HVR-H2 sequence of SEQ ID NO: 67, and an HVR-H3 sequence of SEQ ID NO: 23.
4. The antibody of claim 1 , wherein the antibody comprises an HVR-L1 sequence of SEQ ID NO: 59, an HVR-L2 sequence of SEQ ID NO:3, an HVR-L3 sequence of SEQ ID NO: 4, an HVR-H1 sequence of SEQ ID NO: 11, an HVR-H2 sequence of SEQ ID NO: 67, and an HVR-H3 sequence of SEQ ID NO: 23.
5. The antibody of claim 1 , wherein the antibody comprises a heavy chain variable region comprising a sequence with at least 95% sequence identity to SEQ ID NO. 72 and a light chain variable region comprising a sequence with at least 95% sequence identity to SEQ ID NO: 65.
6. The antibody of claim 5 , wherein the antibody comprises a heavy chain variable region comprising the sequence of SEQ ID NO. 72 and a light chain variable region of SEQ ID NO: 65.
7. (The antibody of claim 1 , comprising light chain and heavy chain amino acid sequences with at least 95% sequence identity to the amino acid sequences of one of the following combinations of sequences: SEQ ID NOs: 5 and 32; SEQ ID NOs: 63 and 32; SEQ ID NOs: 65 and 32; SEQ ID NOs: 5 and 72; SEQ ID NOs: 63 and 72; and SEQ ID NOs: 65 and 72; SEQ ID NOs: 62 and 32; SEQ ID NOs: 64 and 32; SEQ ID NOs: 5 and 70; SEQ ID NOs: 5 and 71; SEQ ID NOs: 66 and 32.
8. The antibody of claim 7 , comprising light chain and heavy chain amino acid sequences of one of the following combinations of sequences: SEQ ID NOs: 5 and 32; SEQ ID NOs: 63 and 32; SEQ ID NOs: 65 and 32; SEQ ID NOs: 5 and 72; SEQ ID NOs: 63 and 72; and SEQ ID NOs: 65 and 72; SEQ ID NOs: 62 and 32; SEQ ID NOs: 64 and 32; SEQ ID NOs: 5 and 70; SEQ ID NOs: 5 and 71; SEQ ID NOs: 66 and 32.
9. The isolated antibody of claim 1 , wherein the antibody specifically binds to a K11-linked polyubiquitinated protein.
10. The isolated antibody of claim 1 , wherein the antibody modulates at least one polyubiquitin-mediated signaling pathway.
11. The isolated antibody of claim 1 , wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody, or an antibody fragment that binds K11-linked polyubiquitin.
12. The antibody of claim 1 , wherein the antibody binds K11-linked polyubiquitin under stringent conditions, but does not bind K48-linked polyubiquitin under stringent conditions, wherein the stringent conditions comprise 4M urea.
13. The antibody of claim 1 , wherein the antibody binds K11-linked polyubiquitin under stringent conditions, but does not bind K63-linked polyubiquitin under stringent conditions, wherein the stringent conditions comprise 4M urea.
14. The antibody of claim 1 , wherein the antibody binds K11-linked polyubiquitin under stringent conditions, but does not bind K48-linked polyubiquitin or K63-linked polyubiquitin under stringent conditions, wherein the stringent conditions comprise 4 M urea.
15. The antibody of claim 1 , wherein the antibody does not bind monoubiquitin.
16. An immunoconjugate comprising the antibody of claim 1 and a cytotoxic agent.
17. A pharmaceutical formulation comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.
18. The pharmaceutical formulation of claim 17 , further comprising an additional therapeutic agent.
19. An isolated monoclonal antibody that binds to the same antigenic determinant on K11-linked ppolyubiquitin as the antibody of claim 6 .
20. An isolated monocianal antibody that competes with the antibody of claim 6 for binding to K11-linked polyubiquitin.