Purification of Common Lung Cancer Antigen recognized with Monoclonal Antibody TFS and Production of Monoclonal Antibodies of Second Generation by its antigen.

点击翻译按钮获取中文摘要

Monoclonal antibody(MAb)TFS-2 was produced against Small Cell Lung Cancer and reacted with all tissue types of lung cancer. Recently we tried the radioimmunodetection of human lung cancer with ^<99m>Tc labeled TFS-2 followed by getting clear imaging of primary and metastatic lung cancers in the various tissues such as bone, lung, and liver.(1)Purification of Common Lung Cancer Antigen.Masses of metastatic small cell lung cancer to liver was used as source for purification. Samples were solubilized by PBS with 0.5% of NP-40. Samples were twice passed through Affinity Chromatography(tresyl-activated Sepharose 4B couppled with MAb TFS-2). NP-40 was removed by ethanol pi-ecipitatiori. SDS-PAGE showed 2 bands of protein, 39kDa and 33kDa respectedly. Band of 33 kDa reacted with MAb TFS-2 on the Western blotting. Protein of 33kDa was analyzed directly transferring to membrane and 20 amino acid residue was determined.(2)Analysis of Gene coding Common Lung Cancer Antigen.The MRNA was obtained from small cell lutig cancer cell line NCI-1169, and the expression CDNA library in plasmid was constructed from the MRNA. The plasmids were transfected to Cos-1 cells and clones expressing antigen on the cell surface were screened by ^<125>I labeled MAb TFS-2. A EDNA clone coding the protein reacted with MAb TFS-2 was isolated.(3)Production of Monoclonal antibodies of the second generation Several clones of hybridoma cell that reacted with purified antigen presently were obtained.