Identification of a novel serum protein secreted by lung carcinoma cells.
The murine anti-human lung tumor monoclonal antibody L3 recognizes antigens found both in the medium of cultured carcinoma cells and in normal human serum. Sequential immunoprecipitation experiments indicate that the L3 antigen is also recognized by a previously described monoclonal antibody directed against a melanoma-associated antigen [Natali, P. G., Wilson, B. S., Imai, K., Bigotti, A., & Ferrone, S. (1982) Cancer Res. 42, 583-589]. This antibody precipitated a Mr 76000 glycoprotein from metabolically labeled extracts of the lung carcinoma cell line Calu-1 and a Mr 94 000 glycoprotein from labeled culture medium. Pulse-chase experiments suggested a precursor-product relationship between these molecules. Analysis of glycosidase sensitivities of the two forms indicated that maturation of carbohydrate side chains correlated with the apparent increase in molecular weights. L3 antigenic activity, measured in a competitive radiometric cell binding assay, was purified more than 90-fold from serum-free medium of Calu-1 cells and more than 3000-fold from normal human serum. The major immunoreactive components purified from culture medium and serum were identical with respect to apparent molecular weight, electrophoretic mobility, pI, glycosidase sensitivity, and V8 protease fingerprints. In addition, the sequence of the amino-terminal 16 N-terminal amino acid residues of the major immunoreactive species from both sources was identical. The properties of the L3 antigen did not correspond to those of any known protein, suggesting that this serum protein has not been previously characterized.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D001798:Blood Proteins / Q000302:isolation & purification*; D002460:Cell Line; D002846:Chromatography, Affinity; D003470:Culture Media; D005455:Fluorescent Antibody Technique; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood* / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D001798:Blood Proteins; D003470:Culture Media