Anti-GPNMB antibodies and diagnostic uses thereof
The present application discloses anti-GPNMB antibodies and methods of using the same. The antibodies are selected to bind to an epitope located in a C-terminal fragment of the protein and are selected for specificity in immunohistochemical assays. The antibodies are useful in, for example, immunohistochemical methods of detecting GPNMB-positive cells in tumors.
1. A monoclonal antibody capable of specifically binding to a C-terminal portion of human transmembrane glycoprotein NMB (GPNMB) comprising amino acids 22-486 of SEQ ID NO: 1, said monoclonal antibody comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises 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; and
(c) an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 4; and
wherein the VL comprises 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 monoclonal antibody of claim 1 , wherein the VH further comprises the following heavy chain variable domain framework regions (FR-H):
(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 monoclonal antibody of claim 1 , wherein the VL further comprises the following light chain variable domain framework regions (FR-L):
(k) FR-L1 comprising the amino acid sequence of SEQ ID NO: 12;
(1) 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 monoclonal antibody of claim 1 , wherein the VH comprises a sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16.
5. The monoclonal antibody of claim 4 , wherein the VH comprises SEQ ID NO: 16.
6. The monoclonal antibody of claim 1 , wherein the heavy chain further comprises the following heavy chain variable domain framework regions (FR-H):
(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; and
wherein the light chain further comprises the following light chain variable domain framework regions (FR-L):
(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.
7. The monoclonal antibody of claim 1 , wherein the antibody is a rabbit antibody.
8. The monoclonal antibody of claim 1 , wherein the monoclonal antibody is an IgG antibody.
9. A method of detecting the presence or expression level of GPNMB in a biological sample comprising contacting the biological sample with the monoclonal antibody of claim 1 and detecting the presence of the bound antibody.
10. The method of claim 9 , wherein the detecting is by immunohistochemistry (IHC), immunofluorescence, or immunoblot.
11. The method of claim 10 , wherein the detecting is by IHC.
12. The method of claim 9 , wherein the sample comprises a fixed tissue.
13. The method of claim 12 , wherein the fixed tissue is a formalin fixed, paraffin embedded (FFPE) tissue.
14. The method of claim 9 , wherein the sample is from a subject having cancer or autoimmune disease.
15. A kit comprising the monoclonal antibody according to claim 1 .
16. The monoclonal antibody of claim 1 , wherein the VL comprises a sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 17.
17. The monoclonal antibody of claim 1 , wherein the VH comprises a sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16, and the VL comprises a sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 17.
18. The monoclonal antibody of claim 16 , wherein the VL comprises SEQ ID NO: 17.