title: Cobra venom factor and human C3 share carbohydrate antigenic determinants.
abstract: As tools to study structural relationships of cobra venom factor (CVF) and human complement component C3, murine monoclonal antibodies to CVF were produced. In this paper we describe two of these monoclonal anti-CVF antibodies designated GV1.8 and GV1.10, both of which bind to carbohydrate epitopes. On immunoblotting, antibody GV1.8 binds to both the alpha- and beta-chains of CVF, whereas antibody GV1.10 binds only to the alpha-chain of CVF. After enzymatic deglycosylation of CVF with N-glycanase (peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase), both antibodies lose their ability to bind to the deglycosylated protein. Additionally, the free oligosaccharide chains of CVF are able to inhibit the binding of antibodies GV1.8 and GV1.10 to CVF on enzyme-linked immunosorbent assay, further demonstrating their carbohydrate specificity. Both monoclonal antibodies to CVF cross-react with human C3. Antibody GV1.8 binds to both chains of human C3 indicating that the shared antigenic epitope present on the two glycosylated chains of CVF is also present on the two chains of human C3. Antibody GV1.10 cross-reacts only with the beta-chain of human C3 which is the homologous chain to the alpha-chain of CVF. After enzymatic deglycosylation of human C3 by N-glycanase, both antibodies lose their ability to bind to the deglycosylated protein consistent with the carbohydrate nature of the recognized epitopes. These results indicate that CVF and human C3 share carbohydrate epitopes on their homologous and nonhomologous chains.
mesh_terms: D002241:Carbohydrates / Q000276:immunology; D003176:Complement C3 / Q000276:immunology*; D003429:Cross Reactions; D004546:Elapid Venoms / Q000276:immunology*; D000939:Epitopes; D006023:Glycoproteins / Q000276:immunology*; D006026:Glycoside Hydrolases; D006801:Humans; D007163:Immunosorbent Techniques
chemical_list: D002241:Carbohydrates; D003176:Complement C3; D004546:Elapid Venoms; D000939:Epitopes; D006023:Glycoproteins; C016310:cobra venom factor; D006026:Glycoside Hydrolases; C020668:glycanase