Carbohydrate epitope defined by an antitumor monoclonal antibody detected on glycoproteins and a glycolipid by immunoblotting.
The murine monoclonal antibody (MAb) MOv2 was found to be directed against the carbohydrate moieties of different kinds of molecules expressed on a human ovarian cystoadenocarcinoma. To define further the glycoconjugates carrying the MOv2-defined epitope, different procedures were used to analyze materials from surgical specimens and carcinoma cell lines. SDS-PAGE and immunoblotting showed glycoprotein molecules migrating in the gel as high and intermediate molecular weight components. A low-molecular-weight band, migrating approximately with the dye front, was also immunostained by MOv2. On the other hand, the immunostaining of high-performance thin-layer chromatography (HPTLC) of the total glycolipid extract and its neutral and acid fractions, after DEAE chromatography, showed selective reactivity with a neutral glycolipid. Reanalysis by immunoblotting of this glycolipid band scraped off the HPTLC plate indicated that it corresponds to the low-molecular-weight component. Periodate oxidation and Pronase digestion further demonstrate the saccharide nature of the determinant on both types of glycoconjugates. In conclusion, we report evidence that with a single analytical procedure, i.e., immunoblotting, it is possible to recognize the same carbohydrate determinant carried on both protein and lipid molecules.
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis*; D002460:Cell Line; D003536:Cystadenocarcinoma / Q000032:analysis*; D000939:Epitopes / Q000032:analysis*; D005260:Female; D006017:Glycolipids / Q000032:analysis* / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D010051:Ovarian Neoplasms / Q000032:analysis*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002241:Carbohydrates; D000939:Epitopes; D006017:Glycolipids; D006023:Glycoproteins