Epitope Mapping Using the Cell-Based 2 × Alanine Substitution Method About the Anti-mouse CXCR6 Monoclonal Antibody, Cx 6 Mab-1.

An anti-mouse CXC chemokine receptor 6 (mCXCR6) monoclonal antibody (mAb), CxMab-1, was developed recently. CxMab-1 is applicable for flow cytometry, Western blotting, and enzyme-linked immunosorbent assay. The purpose of this study is to determine the binding epitope of CxMab-1 using 2 × alanine mutated mCXCR6. Analysis of flow cytometry revealed that CxMab-1 did not recognize S8A-A9G, L10A-Y11A, D12A-G13A, and H14A-Y15A mutants of mCXCR6. The results clearly indicate that the binding epitope of CxMab-1 includes Ser8, Ala9, Leu10, Tyr11, Asp12, Gly13, His14, and Tyr15 of mCXCR6. The successful determination of the CxMab-1 epitope might contribute to the pathophysiological investigation of mCXCR6.