Isolation of small cell lung cancer-associated antigen from human brain.
Previous studies have demonstrated that monoclonal antibody TFS-4 recognizes a cell surface antigen with a molecular weight of 124,000 expressed selectively on small-cell lung cancer but not on non-small-cell lung cancers and that it cross-reacts with human brain. The antigenic determinant on small-cell lung cancer and that on brain shared common characteristics, i.e., trypsin sensitivity, heat lability, and neuraminidase resistance, suggesting that they are similar peptides (T. Okabe et al., Cancer Res., 44: 5273-5278, 1984; J-i. Watanabe et al., Cancer Res., 47:826-829, 1987). In order to elucidate the nature of this unique antigen recognized by TFS-4, we have purified the antigen to homogeneity from human brain. The antigen was solubilized from brain with 0.5% Nonidet P-40, precipitated with 50% ammonium sulfate, and subsequently purified by sequential chromatographies, i.e., diethylaminoethyl-Sepharose ion exchange, immunoaffinity, and gel permeation high-pressure liquid chromatography. The antigenic reactivity was assessed by immunoblotting using TFS-4 as a primary antibody. The purified antigen showed a single protein band with a molecular weight of 124,000 on sodium dodecylsulfate-polyacrylamide gel electrophoresis detected by a silver staining technique. The results suggest that the antigen on brain tissues is structurally related to the molecule expressed on small-cell lung cancer.
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D001923:Brain Chemistry; D018288:Carcinoma, Small Cell / Q000276:immunology*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007163:Immunosorbent Techniques; D008175:Lung Neoplasms / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes