Antibodies directed to the deletion mutants of epidermal growth factor receptor and uses thereof
The present invention relates to novel antibodies, particularly antibodies directed against deletion mutants of epidermal growth factor receptor and particularly to the type III deletion mutant, EGFRvIII. The invention also relates to human monoclonal antibodies directed against deletion mutants of epidermal growth factor receptor and particularly to EGFRvIII. Diagnostic and therapeutic formulations of such antibodies, and immunoconjugates thereof, are also provided.
1. An isolated polynucleotide molecule:
(a) comprising a nucleotide sequence encoding the heavy chain amino acid sequence selected from the group consisting of: SEQ ID NOs: 2, 4, 5, 7, and 10, or encoding a heavy chain amino acid sequence, or a fragment thereof, selected from the group consisting of SEQ ID NOs: 142, 9, 12, 13, 15, 16 and 17, wherein the fragment comprises a CDR1 that is a CDR1 in the heavy chain amino acid sequence, a CDR2 that is a CDR2 in the heavy chain amino acid sequence, and a CDR3 that is a CDR3 in the heavy chain amino acid sequence; or
(b) consisting of a nucleotide sequence encoding a fragment of the heavy chain amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 5, 7 and 10, wherein the fragment comprises a CDR1 that is a CDR1 in the heavy chain amino acid sequence, a CDR2 that is a CDR2 in the heavy chain amino acid sequence, and a CDR3 that is a CDR3 in the heavy chain amino acid sequence.
2. An isolated polynucleotide molecule comprising:
(a) a nucleotide sequence encoding the light chain amino acid sequence selected from the group consisting of SEQ ID NOs: 19, 21, 25, 26, 28, 31, 32, 33, and 144, or encoding a light chain amino acid sequence, or a fragment thereof, selected from the group consisting of SEQ ID NOs: 20, 29, and 23, wherein the fragment comprises a CDR1 that is a CDR1 in the light chain amino acid sequence, a CDR2 that is a CDR2 in the light chain amino acid sequence, and a CDR3 that is a CDR3 in the light chain amino acid sequence; or
(b) consisting of a nucleotide sequence encoding a fragment of the light chain amino acid sequence selected from the group consisting of SEQ ID NOs: 19, 21, 25, 26, 28, 31, 32, 33, and 144, wherein the fragment comprises a CDR1 that is a CDR1 in the light chain amino acid sequence, a CDR2 that is a CDR2 in the light chain amino acid sequence, and a CDR3 that is a CDR3 in the light chain amino acid sequence.
3. The isolated polynucleotide molecule of claim 1 , wherein the heavy chain amino acid sequence comprises SEQ ID NO: 2.
4. The isolated polynucleotide molecule of claim 1 , wherein the heavy chain amino acid sequence comprises SEQ ID NO: 142.
5. The isolated polynucleotide molecule of claim 2 , wherein the light chain amino acid sequence comprises SEQ ID NO: 19.
6. The isolated polynucleotide molecule of claim 2 , wherein the light chain amino acid sequence comprises SEQ ID NO: 144.
7. An isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain polypeptide comprising the following complementarity determining regions (CDRs): a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 2; a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 2; and a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 2.
8. The isolated polynucleotide molecule of claim 7 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
9. The isolated polynucleotide molecule of claim 7 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
10. The isolated polynucleotide molecule of claim 7 , wherein the heavy chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 108, SEQ ID NO: 110, and SEQ ID NO: 112.
11. An isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain polypeptide comprising the following complementarity determining regions (CDRs): a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 142; a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 142; and a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 142.
12. The isolated polynucleotide molecule of claim 11 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
13. The isolated polynucleotide molecule of claim 11 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
14. The isolated polynucleotide molecule of claim 11 , wherein the heavy chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 1-21, SEQ ID NO: 123, and SEQ ID NO: 125.
15. An isolated polynucleotide molecule comprising nucleotide sequence encoding a heavy chain polypeptide comprising a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 142.
16. An isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain polypeptide comprising a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 2.
17. An isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain polypeptide comprising the following complementarity determining regions (CDRs): a light chain CDR1 that is a CDR1 in SEQ ID NO: 19; a light chain CDR2 that a CDR2 in SEQ ID NO: 19; and a light chain CDR3 that is a CDR3 in SEQ ID NO: 19.
18. The isolated polynucleotide molecule of claim 17 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
19. The isolated polynucleotide molecule of claim 17 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
20. The isolated polynucleotide molecule of claim 17 , wherein the light chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 101, SEQ ID NO: 103, and SEQ ID NO: 105.
21. An isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain polypeptide comprising the following complementarity determining regions (CDRs): a light chain CDR1 that is a CDR1 in SEQ ID NO: 144; a light chain CDR2 that a CDR2 in SEQ ID NO: 144; and a light chain CDR3 that is a CDR3 in SEQ ID NO: 144.
22. The isolated polynucleotide molecule of claim 21 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
23. The isolated polynucleotide molecule of claim 21 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
24. The isolated polynucleotide molecule of claim 21 , wherein the light chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 101, SEQ ID NO: 116, and SEQ ID NO: 118.
25. An isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain polypeptide comprising a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19.
26. A composition comprising:
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain polypeptide comprising the following complementarity determining regions (CDRs): a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 2; a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 2; a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 2; and
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain polypeptide comprising the following CDRs: a light chain CDR1 that is a CDR1 in SEQ ID NO: 19; a light chain CDR2 that a CDR2 in SEQ ID NO: 19; and a light chain CDR3 that is a CDR3 in SEQ ID NO: 19,
wherein the heavy chain polypeptide and light chain polypeptide are of a monoclonal antibody that specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
27. The composition of claim 26 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
28. The composition of claim 26 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
29. The composition of claim 26 , wherein the heavy chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 108, SEQ ID NO: 110, and SEQ ID NO: 112, and wherein the light chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 101, SEQ ID NO: 103, and SEQ ID NO: 105.
30. A composition comprising:
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain polypeptide comprising the following complementarity determining regions (CDRs): a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 142; a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 142; a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 142; and
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain polypeptide comprising the following CDRs: a light chain CDR1 that is a CDR1 in SEQ ID NO: 144; a light chain CDR2 that a CDR2 in SEQ ID NO: 144; and a light chain CDR3 that is a CDR3 in SEQ ID NO: 144;
wherein the heavy chain polypeptide and light chain polypeptide are of a monoclonal antibody that specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
31. The composition of claim 30 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
32. The composition of claim 30 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
33. The composition of claim 30 , wherein the heavy chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 121, SEQ ID NO: 123, and SEQ ID NO: 125, and wherein the light chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 101, SEQ ID NO: 116, and SEQ ID NO: 118.
34. A composition comprising:
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain polypeptide comprises a variable region that is at least 90% identical to the amino acid sequence selected from the group consisting of: SEQ ID NO: 2 and SEQ ID NO: 142; and
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain polypeptide comprising a variable region that is at least 90% identical to the amino acid sequence selected from the group consisting of: SEQ ID NO: 19 and SEQ ID NO: 144.
35. The composition of claim 34 , wherein the heavy chain polypeptide comprises a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 2 and the light chain polypeptide comprises a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19.
36. The composition of claim 34 , wherein the heavy chain polypeptide comprises a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 142 and the light chain polypeptide comprises a variable region that is at least 95% identical to the amino acid sequence of SEQ ID NO: 144.
37. An isolated polynucleotide comprising a nucleotide sequence encoding a monoclonal antibody that binds to EGFRvIII, wherein the monoclonal antibody comprises a light chain polypeptide and a heavy chain polypeptide,
wherein the heavy chain polypeptide comprises following complementarity determining regions (CDRs): a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 2; a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 2; a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 2; and
wherein the light chain polypeptide comprises the following CDRs: a light chain CDR1 that is a CDR1 in SEQ ID NO: 19; a light chain CDR2 that a CDR2 in SEQ ID NO: 19; and a light chain CDR3 that is a CDR3 in SEQ ID NO: 19,
wherein the monoclonal antibody specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
38. The isolated polynucleotide of claim 37 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
39. The isolated polynucleotide of claim 37 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
40. The isolated polynucleotide of claim 37 , wherein the heavy chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 108, SEQ ID NO: 110, and SEQ ID NO: 112, and wherein the light chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 101, SEQ ID NO: 103, and SEQ ID NO: 105.
41. An isolated polynucleotide comprising a nucleotide sequence encoding a monoclonal antibody that binds to EGFRvIII, wherein the monoclonal antibody comprises a light chain polypeptide and a heavy chain polypeptide,
wherein the heavy chain polypeptide comprises following complementarity determining regions (CDRs): a heavy chain CDR1 that is a CDR1 in SEQ ID NO: 142; a heavy chain CDR2 that is a CDR2 in SEQ ID NO: 142; a heavy chain CDR3 that is a CDR3 in SEQ ID NO: 142; and
wherein the light chain polypeptide comprises the following CDRs: a light chain CDR1 that is a CDR1 in SEQ ID NO: 144; a light chain CDR2 that a CDR2 in SEQ ID NO: 144; and a light chain CDR3 that is a CDR3 in SEQ ID NO: 144,
wherein the monoclonal antibody specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
42. The isolated polynucleotide of claim 41 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
43. The isolated polynucleotide of claim 41 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
44. The isolated polynucleotide molecule of claim 41 , wherein the heavy chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 121, SEQ ID NO: 123, and SEQ ID NO: 125, and wherein the light chain polypeptide comprises all three of the following amino acid sequences: SEQ ID NO: 101, SEQ ID NO: 116, and SEQ ID NO: 118.
45. An isolated polynucleotide comprising a nucleotide sequence encoding a monoclonal antibody that binds to EGFRvIII, wherein the monoclonal antibody comprises a light chain variable region and a heavy chain variable region,
wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 2, and
wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 19.
46. An isolated polynucleotide comprising a nucleotide sequence encoding a monoclonal antibody that binds to EGFRvIII, wherein the monoclonal antibody comprises a light chain variable region and a heavy chain variable region,
wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 142, and
wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 144.
47. A composition comprising:
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a heavy chain amino acid sequence, or a fragment thereof, selected from the group consisting of SEQ ID NOs: 142, 2, 4, 5, 7, 9, 10, 12, 13, 15, 16 and 17; and
an isolated polynucleotide molecule comprising a nucleotide sequence encoding a light chain amino acid sequence, or a fragment thereof, selected from the group consisting of SEQ ID NOs: 144, 19, 20, 21, 29, 23, 25, 26, 28, 33, 31 and 32.
48. The isolated polynucleotide molecule of claim 1 , encoding a binding fragment of an antibody.
49. The isolated polynucleotide molecule of claim 48 , wherein the binding fragment comprises an entire variable region.
50. The isolated polynucleotide molecule of claim 49 , wherein the isolated polynucleotide molecule encodes a sequence selected from the group consisting of: SEQ ID NOs: 142, 9, 12, 13,15, 16, 17, 2, 4, 5, 7 and 10.
51. The isolated polynucleotide molecule of claim 49 , wherein the heavy chain amino acid sequence is of an antibody that specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
52. The isolated polynucleotide molecule of claim 1 , wherein the heavy chain amino acid sequence is of an antibody that specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
53. The isolated polynucleotide molecule of claim 2 , encoding a binding fragment of an antibody.
54. The isolated polynucleotide molecule of claim 53 , wherein the binding fragment comprises an entire variable region.
55. The isolated polynucleotide molecule of claim 54 , wherein the isolated polynucleotide molecule encodes a sequence selected from the group consisting of: SEQ ID NOs: 19, 21, 25, 26, 28, 31, 32, 33, 144, 20, 29, and 23.
56. The isolated polynucleotide molecule of claim 54 , wherein the light chain amino acid sequence is of an antibody that specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
57. The isolated polynucleotide molecule of claim 2 , wherein the light chain amino acid sequence is of an antibody that specifically binds to a peptide comprising EEKKGNYVVT (SEQ ID NO: 94).
58. The isolated polynucleotide molecule of claim 1 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
59. The isolated polynucleotide molecule of claim 1 , wherein each CDR is defined in accordance with the CDR definition of Chothia.
60. The isolated polynucleotide molecule of claim 2 , wherein each CDR is defined in accordance with the CDR definition of Kabat.
61. The isolated polynucleotide molecule of claim 2 , wherein each CDR is defined in accordance with the CDR definition of Chothia.