[Preparation and application of mouse monoclonal antibodies against human cyclophilin A (CypA)]

Objective To prepare monoclonal antibodies against human cyclophilin A (CypA) and explore the role of CypA in the occurrence and development of colon cancer. Methods According to the CypA gene sequence, the recombinant protein of CypA was expressed by the prokaryotic expression vector and purified by a nickel affinity chromatography column. The monoclonal antibody was prepared by the antigen which is purified protein immunizing mice and hybridoma technique. Utilizing CypA monoclonal antibody , immunohistochemistry was used to detect the difference expression between colorectal cancer and adjacent tissues, meanwhile, analyse clinicopathological parameters. Results The prokaryotic expression vector of CypA was successfully constructed, so we obtain the purified protein; One hybridoma cell line secreting anti-CypA antibody was obtained. Immunohistochemical results showed that CypA expressed high level in colorectal cancer and relate with differentiation degree and lymph node metastasis. Conclusion The monoclonal antibody against human CypA was successfully produced in mice, and CypA was overexpressed in colorectal cancer tissues, which was positively correlated with low differentiation and lymph node metastasis.