Epitope specificities and quantitative and serologic aspects of monoclonal complement (C3c and C3d) antibodies.
Twenty-two monoclonal antibodies to human C3c and ten to C3d were obtained by hybridization after the immunization of mice with complement-coated human red cells and/or purified human complement components. C3c antibodies were variable in their agglutination reactions with cells coated with C3 by antibody in vitro; more consistent and potent reactions with these cells were observed with anti-C3d, and all anti-C3d reacted with red cells coated with C3 in vivo. Immunoradiometric assays were used to estimate antibody concentration, affinity, and epitope specificity. The antibody content in ascitic fluids varied from less than 0.1 mg per ml to 5.6 mg per ml. The estimated values of antibody affinities for Sepharose-coupled C3 ranged from 2.8 X 10(6) l per M to 5.0 X 10(8) l per M; on average, IgM antibodies had higher affinities than IgG antibodies. Competitive binding assays showed that the monoclonal antibodies recognized at least seven different epitopes, four on the C3c and three on the C3d fragment of C3. When the results of serologic and quantitative assays were compared, no convincing relationship was found between serologic performance and epitope specificity, antibody concentration, or affinity. IgM antibodies generally gave higher agglutination scores than IgG antibodies, and Ig class was the only useful predictor of serologic efficacy.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000145:classification / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001202:Ascitic Fluid / Q000276:immunology; D003176:Complement C3 / Q000276:immunology*; D015932:Complement C3c; D015933:Complement C3d; D000939:Epitopes; D051379:Mice; D008815:Mice, Inbred Strains; D009994:Osmolar Concentration
D000911:Antibodies, Monoclonal; D003176:Complement C3; D000939:Epitopes; D015932:Complement C3c; D015933:Complement C3d