Anti-Notch3 antibody
Monoclonal antibodies that bind and inhibit activation of human Notch3 are disclosed. The antibodies can be used to treat cell proliferative diseases and disorders, including certain forms of cancer, associated with activation and/or overexpression of Notch3.
1. An isolated antibody that binds human Notch3 comprising:
(i) an immunoglobulin heavy chain variable region comprising a CDR H1 comprising the amino acid sequence of SEQ ID NO: 3, a CDR H2 comprising the amino acid sequence of SEQ ID NO: 4, and a CDR H3 comprising the amino acid sequence of SEQ ID NO: 5; and
(ii) an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 22, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9.
2. The isolated antibody of claim 1 , wherein the immunoglobulin light chain variable region is selected from:
(a) an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9;
(b) an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9;
(c) an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 22, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9; and
(d) an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 22, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9.
3. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin heavy chain variable region of claim 1 .
4. An expression vector comprising the nucleic acid of claim 3 .
5. A host cell comprising the expression vector of claim 4 .
6. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin light chain variable region of claim 1 .
7. An expression vector comprising the nucleic acid of claim 6 .
8. A host cell comprising the expression vector of claim 7 .
9. The antibody of claim 1 , wherein the antibody has a K D of 15 nM or lower, as measured by surface plasmon resonance or bio-layer interferometry.
10. A method of inhibiting proliferation of a tumor cell that overexpresses Notch3 comprising exposing the cell to an effective amount of the antibody of claim 1 to inhibit proliferation of the tumor cell.
11. A method of inhibiting growth of a tumor that overexpresses Notch3 in a mammal, the method comprising exposing the mammal to an effective amount of the antibody of claim 1 to inhibit growth of the tumor.
12. A method of treating cancer that overexpresses Notch3 in a mammal, the method comprising administering an effective amount of the antibody of claim 1 to a mammal in need thereof.
13. The isolated antibody of claim 1 , wherein the immunoglobulin light chain variable region comprises a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8, and a CDRL 3 comprising the amino acid sequence of SEQ ID NO: 9.
14. The isolated antibody of claim 1 , wherein the immunoglobulin light chain variable region comprises a CDR L1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9.
15. The isolated antibody of claim 1 , wherein the immunoglobulin light chain variable region comprises a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 22, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9.
16. The isolated antibody of claim 1 , wherein the immunoglobulin light chain variable region comprises a CDR L1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 22 , and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9.
17. An isolated antibody that binds human Notch3, comprising an immunoglobulin heavy chain variable region comprising the amino acid sequence of SEQ ID NO:6, and an immunoglobulin light chain variable region comprising the amino acid sequence of SEQ ID NO:10, 23, 24, 25, 26, 27, 28, 29, 37, 38, or 39.
18. An isolated nucleic acid comprising a nucleotide sequence encoding the immunoglobulin heavy chain variable region of claim 17 .
19. An expression vector comprising the nucleic acid of claim 18 .
20. A host cell comprising the expression vector of claim 19 .
21. An isolated nucleic acid comprising a nucleotide sequence encoding the immunoglobulin light chain variable region of claim 17 .
22. An expression vector comprising the nucleic acid of claim 21 .
23. A host cell comprising the expression vector of claim 22 .
24. The isolated antibody of claim 17 , wherein the immunoglobulin light chain variable region comprises the amino acid sequence of SEQ ID NO:10.
25. The isolated antibody of claim 17 , wherein the immunoglobulin light chain variable region comprises the amino acid sequence of SEQ ID NO:23.
26. An isolated antibody that binds human Notch3 comprising an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 1, and an immunoglobulin light chain comprising the amino acid sequence selected from SEQ ID NO: 2, 30, 31, 32, 33, 34, 35, 36, 40, 41, or 42.
27. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin heavy chain of claim 26 .
28. An expression vector comprising the nucleic acid of claim 27 .
29. A host cell comprising the expression vector of claim 28 .
30. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin light chain of claim 26 .
31. An expression vector comprising the nucleic acid of claim 30 .
32. A host cell comprising the expression vector of claim 31 .
33. The isolated antibody of claim 26 , wherein the immunoglobulin light chain comprises the amino acid sequence of SEQ ID NO:2.
34. The isolated antibody of claim 26 , wherein the immunoglobulin light chain comprises the amino acid sequence of SEQ ID NO:30.
35. A method of producing an antibody that binds human Notch3 or an antigen binding fragment of the antibody, the method comprising:
(a) growing a host cell comprising:
(i) an expression vector comprising a nucleic acid encoding
an immunoglobulin heavy chain variable region comprising a CDR H1 comprising the amino acid sequence of SEQ ID NO: 3, a CDR H2 comprising the amino acid sequence of SEQ ID NO: 4, and a CDR H3 comprising the amino acid sequence of SEQ ID NO: 5; and
an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 22, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9; or
(ii) an expression vector comprising a nucleic acid encoding an immunoglobulin heavy chain variable region comprising a CDR H1 comprising the amino acid sequence of SEQ ID NO: 3, a CDR H2 comprising the amino acid sequence of SEQ ID NO: 4, and a CDR H3 comprising the amino acid sequence of SEQ ID NO: 5; and
an expression vector encoding an immunoglobulin light chain variable region comprising a CDR L1 comprising the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 21, a CDR L2 comprising the amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 22, and a CDR L3 comprising the amino acid sequence of SEQ ID NO: 9;
under conditions so that the host cell expresses a polypeptide comprising the immunoglobulin heavy chain variable region and the immunoglobin light chain variable region, thereby producing the antibody or the antigen-binding fragment of the antibody; and
(b) purifying the antibody or the antigen-binding fragment of the antibody.