Preparation Of Monoclonal Antibody Against Antigen Associated Human Lung Cancer And The Immunoaffinity Chromatography Purification Of Its Antigen

Objective To produce the monoclonal antibody (mcAb) specifically reacting with human lung cancer and purificate its antigen from cultured lung cancer cells A549. Methods The mice were immunized with human lung cancer cells A549, the mcAb was screened by indirect cell enzyme-linked immunosorbant assay (ELISA) and immunohistochemistry (IHC), and the hybridoma cell's chromosome was analysed by colchicine; the subclass of the mcAb was identified with Mouse Monoclonal Antibody Isotyping Reagents, and the potency of the mcAb was detected. The mcAb against human lung tumor-associated antigen was coupled with CNBr-activated Sepharose 4B as the ligand for the immunoaffinity chromatography (IAC) to purify the lung tumor-associated antigen from the lysate of lung cancer cells A549. The purified antigen of the mcAb was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western-blot. Results A mcAb named 2B9 was obtained, the chromosome number of the hybridoma cell 2B9 was about 102, the subclass of mcAb was IgM, the mcAb 2B9 can react to lung cancer but very little or not to normal lung tissue and other tumor tissues. The antigen of mcAb 2B9 was purificated by immunoaffinity chromatography from the A549 cell lysate. It has a molecular mass (Mr) of 38Kda. Conclusion A mcAb which can react to lung cancer have been obtained and its antigen was purificated, the antigen of mcAb 2B9 can be a valuable candidate of tumor marker about lung cancer and they may be useful on clinic for diagnosis and prognosis of lung cancer.