Anti-OX40 antibodies and diagnostic uses thereof
The invention provides antibodies immunoreactive with human OX40 and methods of using the same. The antibodies are reactive with a portion of the C-terminus of the human OX40 protein that includes amino acids 266-277. The antibodies are useful for detecting OX40 protein expression in human tissue samples, including by immunohistochemistry, immunofluorescence, or immunoblot.
1. An isolated antibody capable of specifically binding to a C-terminal portion of OX40 comprising amino acids 266-277 of SEQ ID NO: 1, wherein the antibody comprises a heavy chain variable domain (VH) sequence comprising the following heavy chain hypervariable regions (HVR-H):
(a) an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 2;
(b) an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 3;
(c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 4; and and a light chain variable domain (VL) sequence comprising the following light chain hypervariable regions (HVR-L):
(d) an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 9;
(e) an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 10; and
(f) an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 11.
2. The antibody of claim 1 , wherein the VH sequence further comprises the following heavy chain variable domain framework regions (FR-Hs):
(g) FR-H1 comprising the amino acid sequence of SEQ ID NO: 5;
(h) FR-H2 comprising the amino acid sequence of SEQ ID NO: 6;
(i) FR-H3 comprising the amino acid sequence of SEQ ID NO: 7; and
(j) FR-H4 comprising the amino acid sequence of SEQ ID NO: 8.
3. The antibody of claim 1 , wherein the VL sequence further comprises the following light chain variable domain FRs:
(k) FR-L1 comprising the amino acid sequence of SEQ ID NO: 12;
(l) FR-L2 comprising the amino acid sequence of SEQ ID NO: 13;
(m) FR-L3 comprising the amino acid sequence of SEQ ID NO: 14; and
(n) FR-L4 comprising the amino acid sequence of SEQ ID NO: 15.
4. The antibody of claim 1 , wherein:
the VH sequence has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16; or
the VL sequence has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 17; or
the VH sequence has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16 and the VL sequence has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 17.
5. The antibody of claim 1 , wherein the VH sequence comprises SEQ ID NO: 16.
6. The antibody of claim 1 , wherein the VL sequence comprises SEQ ID NO: 17.
7. The antibody of claim 1 , wherein the antibody further comprises the following heavy chain variable domain framework regions (FR-Hs) and light chain variable domain framework regions (FR-Ls):
(g) FR-H1 comprising the amino acid sequence of SEQ ID NO: 5;
(h) FR-H2 comprising the amino acid sequence of SEQ ID NO: 6;
(i) FR-H3 comprising the amino acid sequence of SEQ ID NO: 7;
(j) FR-H4 comprising the amino acid sequence of SEQ ID NO: 8;
(k) FR-L1 comprising the amino acid sequence of SEQ ID NO: 12;
(l) FR-L2 comprising the amino acid sequence of SEQ ID NO: 13;
(m) FR-L3 comprising the amino acid sequence of SEQ ID NO: 14; and
(n) FR-L4 comprising the amino acid sequence of SEQ ID NO: 15.
8. The antibody of claim 1 , wherein the antibody comprises a VH sequence of SEQ ID NO: 16 and a VL sequence of SEQ ID NO: 17.
9. The antibody claim 1 , wherein the antibody is a monoclonal antibody.
10. The antibody of claim 9 , wherein the monoclonal antibody is a rabbit monoclonal antibody.
11. The antibody of claim 1 , wherein the antibody is an IgG antibody.
12. The antibody of claim 1 , wherein the antibody is an antibody fragment that specifically binds OX40.
13. The antibody of claim 12 , wherein the antibody fragment is selected from the group consisting of Fab, single chain variable fragment (scFv), Fv, Fab′, Fab′-SH, F(ab′)2, and diabody.
14. A kit comprising an antibody according to claim 1 .
15. The kit of claim 14 , further comprising a secondary antibody immunoreactive with the antibody capable of specifically binding to the C-terminal portion of OX40.
16. The kit of claim 15 , further comprising an enzyme coupled to or adapted to be coupled to the secondary antibody.
17. The kit of claim 16 , further comprising a set of reagents reactive with the enzyme to deposit a chromogen or fluorophore.
18. The kit of claim 17 , wherein the set of reagents reactive with the enzyme comprises a tyramide conjugate.
19. A method of detecting the presence or expression level of OX40 in a biological sample comprising contacting the biological sample with the antibody of claim 1 and detecting the presence of the bound antibody.
20. The method of claim 19 , wherein the detecting is by immunohistochemistry (IHC), immunofluorescence, or immunoblot.
21. The method of claim 20 , wherein the detecting is by IHC.
22. The method of claim 19 , wherein the sample comprises a fixed tissue.
23. The method of claim 22 , wherein the fixed tissue is a FFPE tissue.
24. The method of claim 19 , wherein the sample is from a subject having or predisposed to cancer or autoimmune disease.