Antibodies, compositions, and immunohistochemistry methods for detecting C4.4a
Antibodies, compositions, systems, and methods for detecting C4.4a, for example immunohistochemistry methods for detecting C4.4a using a C4.4a antibody. The antibody may be obtained by immunizing a host with a C4.4a protein such as a peptide downstream of the signal peptide. The antibodies may be adapted to detect the uPAR-like domain 1 and uPAR-like domain 2. Also featured are methods for diagnosing C4.4a-associated tumors using C4.4a antibodies disclosed herein.
1. A monoclonal C4.4a antibody specific for an epitope within a uPAR-like domain 2 within a human C4.4a protein, said monoclonal C4.4a antibody comprising a heavy chain variable region comprising heavy chain complementarity determining regions H-CDR1, H-CDR2, and H-CDR3, wherein H-CDR1 comprises SEQ ID NO: 23, H-CDR2 comprises SEQ ID NO: 25, and H-CDR3 comprises SEQ ID NO: 27, and further comprising a light chain variable region comprising light chain complementarity determining regions L-CDR1, L-CDR2, and L-CDR3, wherein L-CDR1 comprises SEQ ID NO: 31, L-CDR2 comprises SEQ ID NO: 33, and L-CDR3 comprises SEQ ID NO: 35.
2. The monoclonal C4.4a antibody of claim 1 , wherein the heavy chain variable region comprises SEQ ID NO: 17.
3. The monoclonal C4.4a antibody of claim 1 , wherein the uPAR like domain 2 comprises SEQ ID NO: 4.
4. The monoclonal C4.4a antibody of claim 1 , wherein the heavy chain variable region further comprises heavy chain framework regions H-FR1, H-FR2, H-FR3, and H-FR4, wherein H-FR1 comprises SEQ ID NO: 22, H-FR2 comprises SEQ ID NO: 24, H-FR3 comprises SEQ ID NO: 26, and H-FR4 comprises SEQ ID NO: 28.
5. The monoclonal C4.4a antibody of claim 1 , wherein the light chain variable region further comprises light chain framework regions L-FR1, L-FR2, L-FR3, and L-FR4, wherein L-FR1 comprises SEQ ID NO: 30, L-FR2 comprises SEQ ID NO: 32, L-FR3 comprises SEQ ID NO: 34, and L-FR4 comprises SEQ ID NO: 36.
6. The monoclonal C4.4a antibody of claim 1 , wherein the light chain variable region comprises SEQ ID NO: 18.
7. The monoclonal C4.4a antibody of claim 2 , wherein the light chain variable region comprises SEQ ID NO: 18.
8. A kit comprising a C4.4a antibody according to claim 1 .
9. The kit of claim 8 , wherein the monoclonal C4.4a antibody is adapted for immunohistochemistry.
10. The kit of claim 8 further comprising a detection system for making the C4.4a antibody visible.
11. The kit of claim 10 , wherein the detection system comprises a chromogenic detection system.
12. The kit of claim 10 , wherein the detection system comprises a fluorescence detection system.
13. A method of labeling a biological sample for human C4.4a protein, said method comprising:
a. contacting a sample with a C4.4a antibody according to claim 1 ; and
b. making the antibody bound to the sample visible.
14. The method of claim 13 , wherein the biological sample is a tissue sample.
15. The method of claim 14 , wherein the tissue sample is a formalin fixed, paraffin embedded (FFPE) tissue sample.
16. The method of claim 13 , wherein the step of making the antibody visible comprises contacting the antibody with a chromogenic detection system or a fluorescence detection system.
17. The method of claim 13 , wherein the biological sample is from a human.
18. The method of claim 17 , wherein the biological sample is a tissue sample.
19. The method of claim 18 , wherein the tissue sample is a formalin fixed, paraffin embedded (FFPE) tissue sample of a human tumor.
20. A method of diagnosing a C4.4a-associated tumor, said method comprising obtaining a tumor sample, labeling the tumor sample for human C4.4a protein according to the method of claim 13 , and detecting the presence or absence of visible antibody, wherein detection of visible C4.4a antibody is indicative that the tumor is the C4.4a-associated tumor.