Purification Of Human Lung Cancer-associated Antigen

Objective To purify lung cancer-associated antigen N35(N35Ag) by immunoaffinity chromatography using our self-prepared monoclonal antibody N-35(raAb N-35).Method 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 N35Ag by passing the lysate of adenocarcinoma of lung GLC-82 through the prepared column. And additional work was made to identify N35Ag by SDS-PAGE.Result Using the mAb N-35 immunoaffinity column, we got purified N35Ag with a molecular weight of 80KD from the lysate of GLC-82. SDS-PAGE experiment showed that the purified N35Ag has high purity.Conclusion It was high-performance and high-purity that purified raAb 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 was different from them...