A monoclonal antibody against human FXYD6

Previous research has shown that FXYD6 (FXYD domain-containing ion transport regulator 6) is highly increased in bile duct tumor. However, the biological function of FXYD6 is unclear. We aim to prepare and identify a monoclonal antibody against FXYD6, which will be used in diagnostics and as a tool in understanding the role of FXYD6 in pathogenesis of hepatobiliary cancer. In this study, hybridoma cell fusion technology is used for production of FXYD6 monoclonal antibody. BALB/c mice are immunized with FXYD6 synthetic peptide fragment. Hybridoma clones are screened using indirect enzyme-linked immunosorbent assay (ELISA). FXYD6 monoclonal antibody is produced by ascites revulsion. Protein A affinity chromatography is used for the purification of FXYD6 monoclonal antibody. Titer and specificity of monoclonal antibody are assessed by ELISA. Expression of FXYD6 in pancreatic cancer is detected by immunohistochemistry. As a result, one stable hybridoma cell clone producing FXYD6 monoclonal antibody has been established. 2.86 mg monoclonal antibody against FXYD6 with high specificity is prepared with titer of 1:5400. Immunohistochemistry shows that FXYD6 positive staining occurs in the cell membrane of pancreatic cancer, which results in an advantage in investigating the tissue distribution and biological function of FXYD6.