Characterization of the CA 125 antigen associated with human epithelial ovarian carcinomas.
The murine monoclonal antibody OC125 reacts with an antigenic determinant (CA 125) found on a high-molecular-weight glycoprotein complex present in the serum of greater than 80% of women with epithelial ovarian cancer. The antigen expressing CA 125 (CA 125 antigen) isolated from the sera of ovarian carcinoma patients was shown by gel electrophoresis, molecular size exclusion chromatography, and buoyant density ultracentrifugation to have similar immunological and physical characteristics to antigen isolated from an ovarian cancer cell line (OVCA 433) and human milk. A composite sodium dodecyl sulfate: polyacrylamide:1.0% agarose gel resolved the CA 125 activity from the three sources of antigen into disperse bands of similar electrophoretic mobilities with apparent masses of 200,000 to 1 million daltons. The buoyant densities of the CA 125 antigen complexes from human serum, OVCA 433 cells, and human milk were in the range of 1.36 to 1.46 g/ml. Isolation of CA 125 antigen of higher purity from OVCA 433 supernatant was achieved by a series of steps including OC125 immunoaffinity chromatography. Subsequent resolution of this purified CA 125 antigen complex by sodium dodecyl sulfate:polyacrylamide gel electrophoresis gave rise to a band at approximately 200,000 daltons. Treatment of the CA 125 antigen from OVCA 433 cells with 10 mM periodic acid resulted in no loss of activity. Reduction and alkylation in 6 M guanidine-HCl or treatment at 100 degrees C for 20 min resulted in complete loss of activity. Exoglycosidase treatments did not result in loss of activity, whereas protease digestion eradicated all activity. These data strongly suggest that the CA 125 antigenic determinant is composed of, at least in part, conformationally dependent peptide.
D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D015295:Antigens, Tumor-Associated, Carbohydrate; D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity; D000939:Epitopes / Q000032:analysis*; D005260:Female; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006801:Humans; D008970:Molecular Weight; D010051:Ovarian Neoplasms / Q000276:immunology*
D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D002241:Carbohydrates; D000939:Epitopes; D006026:Glycoside Hydrolases