Purification Of Monoclonal Antibody N-35 And Human Lung Cancer-associated Antigen N35

Objective.: To purify mAb N-35 by affinity chromatography using Protein A-Sepharose CL-4B,and to purify lung cancer-associated antigen N35 by immunoaffinity chromatography using our self-prepared monoclonal antibody N-35(mAb N-35).Methods: After culturing P35 hybridoma cells and collecting their culture supernatant, purified mAb N-35 from the supernatant by affinity chromatography using Protein A-Sepharose CL-4B. Then made the mAb N-35 immunoaffinity column using CNBr-antivated Protein A-Sepharose CL-4B and purified mAb N-35.At last,purified antigen N35 by passing the lysate of adenocarcinoma of lung GLC-82 through the prepared column.And additional work was made to identify antigen N35 by SDS-PAGE and Western blot.Results: Using the mAb N-35 immunoaffinity column,we got purified antigen N35 with a molecular weight of 80KD from the lysate of GLC-82.SDS-PAGE experiment showed that the purified antigen N35 has high purity.Conclusion: It was high-performance and high-purity that purified mAb N-35 using Protein A-Sepharose CL-4B. It was high-performance and high-purity that purified N35Ag by immunoaffinity chromatography using CNBr-antivated Protein A-Sepharose CL-4B. N35Ag is different from them in molecular weight and may be a new lung cancer-associated antigen.