Antibodies specific for epidermal growth factor receptor variant III and their uses
The present invention provides antibodies that specifically bind to EGFRvIII (Epidermal Growth Factor Receptor Variant III). The invention further provides bispecific antibodies that bind to EGFRvIII and another antigen (e.g., CD3) as well as antibody conjugates (e.g., antibody-drug-conjugates). The invention further relates to antibody encoding nucleic acids, and methods of obtaining such antibodies (monospecific and bispecific) and antibody conjugates. The invention further relates to therapeutic methods for use of these antibodies and antibody conjugates for the treatment of EGFRvIII-mediated pathologies, including cancer such as glioblastoma.
1. An isolated antibody, which specifically binds to Epidermal Growth Factor Receptor Variant III (EGFRvIII), wherein the antibody comprises
a) a heavy chain variable (VH) region comprising a sequence selected from SEQ ID NOs: 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 25, 30, 32, 34, 35, 37, 39, 41, 43, 44, and 46; and
b) a light chain variable (VL) region comprising a sequence selected from SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 29, 31, 33, 31, 36, 38, 40, 42, 40, 45, and 47.
2. The antibody of claim 1 , wherein the antibody comprises an acyl donor glutamine-containing tag engineered at a specific site.
3. A nucleic acid encoding the antibody of claim 1 .
4. A vector comprising the nucleic acid of claim 3 .
5. A host cell comprising the nucleic acid of claim 3 .
6. A conjugate of the antibody of claim 1 , wherein the antibody is conjugated to an agent, wherein the agent is selected from the group consisting of a cytotoxic agent, an immunomodulating agent, an imaging agent, a therapeutic protein, a biopolymer, and an oligonucleotide.
7. A method of treating a subject in need thereof comprising:
a) providing the antibody according to claim 1 ; and
b) administering said antibody to said patient.
8. A pharmaceutical composition comprising the antibody of claim 1 .
9. A method of treating a condition associated with malignant cells expressing EGFRvIII in a subject comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition of claim 8 .
10. The method of claim 9 , wherein the condition is a cancer.
11. The method of claim 10 , wherein the cancer is an EGFRvIII related cancer selected from the group consisting of glioblastoma multiform, anaplastic astrocytoma, giant cell glioblastoma, gliosarcoma, anaplastic oligodendroglioma, anaplastic ependymoma, anaplastic oligoastrocytoma, choroid plexus carcinoma, anaplastic ganglioglioma, pineoblastoma, pineocytoma, meningioma, medulloepithelioma, ependymoblastoma, medulloblastoma, supraentorial primitive neuroectodermal tumor, atypical teratoid/rhabdoid tumor, mixed glioma, head and neck cancer, non-small cell lung cancer, breast cancer, ovarian cancer, prostate cancer, colorectal cancer, anal cancer, cervical cancer, renal cancer, skin cancer, pancreatic cancer, liver cancer, bladder cancer, gastric cancer, thyroid cancer, mesothelioma, uterine cancer, lymphoma, and leukemia.
12. A method of inhibiting tumor growth or progression in a subject who has malignant cells expressing EGFRvIII, comprising administering to the subject in need thereof an effective amount of the pharmaceutical composition of claim 8 to the subject.
13. A method of inhibiting metastasis of malignant cells expressing EGFRvIII in a subject, comprising administering to the subject in need thereof an effective amount of the pharmaceutical composition of claim 8 to the subject.
14. A method of inducing tumor regression in a subject who has malignant cells expressing EGFRvIII, comprising administering to the subject in need thereof an effective amount of the pharmaceutical composition of claim 8 to the subject.
15. A method of producing an antibody, comprising culturing the host cell of claim 5 under conditions that result in production of the antibody, and isolating the antibody from the host cell or culture.
16. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 9 and a VL region comprising the amino acid sequence of SEQ ID NO: 10.
17. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 13 and a VL region comprising the amino acid sequence of SEQ ID NO: 14.
18. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 25 and a VL region comprising the amino acid sequence of SEQ ID NO: 29.
19. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 39 and a VL region comprising the amino acid sequence of SEQ ID NO: 40.
20. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 46 and a VL region comprising the amino acid sequence of SEQ ID NO: 47.
21. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 15 and a VL region comprising the amino acid sequence of SEQ ID NO: 16.
22. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 41 and a VL region comprising the amino acid sequence of SEQ ID NO: 42.
23. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 11 and a VL region comprising the amino acid sequence of SEQ ID NO: 12.
24. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 37 and a VL region comprising the amino acid sequence of SEQ ID NO: 38.
25. The isolated antibody of claim 1 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 30 and a VL region comprising the amino acid sequence of SEQ ID NO: 31.
26. A nucleic acid encoding the antibody of claim 16 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 9 and a VL region comprising the amino acid sequence of SEQ ID NO: 10.
27. A nucleic acid encoding the antibody of claim 17 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 13 and a VL region comprising the amino acid sequence of SEQ ID NO: 14.
28. A nucleic acid encoding the antibody of claim 18 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 25 and a VL region comprising the amino acid sequence of SEQ ID NO: 29.
29. A nucleic acid encoding the antibody of claim 19 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 39 and a VL region comprising the amino acid sequence of SEQ ID NO: 40.
30. A nucleic acid encoding the antibody of claim 20 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 46 and a VL region comprising the amino acid sequence of SEQ ID NO: 47.
31. A nucleic acid encoding the antibody of claim 21 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 15 and a VL region comprising the amino acid sequence of SEQ ID NO: 16.
32. A nucleic acid encoding the antibody of claim 22 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 41 and a VL region comprising the amino acid sequence of SEQ ID NO: 42.
33. A nucleic acid encoding the antibody of claim 23 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 11 and a VL region comprising the amino acid sequence of SEQ ID NO: 12.
34. A nucleic acid encoding the antibody of claim 24 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 37 and a VL region comprising the amino acid sequence of SEQ ID NO: 38.
35. A nucleic acid encoding the antibody of claim 25 , wherein the antibody comprises a VH region comprising the amino acid sequence of SEQ ID NO: 30 and a VL region comprising the amino acid sequence of SEQ ID NO: 31.