Use of an enzyme-linked immunosorbent assay for screening hybridoma antibodies against hepatitis B surface antigen.
An enzyme-linked immunosorbent assay (ELISA) was developed for screening production of monoclonal antibodies with specificity for HBsAg. Mouse hybridoma IgG were firstly extracted from assay medium with goat anti-mouse IgG adsorbed on polystyrene beads. The specific antibody was revealed by saturation with HBs antigen from human positive sera followed by reaction with specific sheep anti-HBs antibody conjugated with peroxidase. The sensitivity was of the order of 0.5-1 ng/ml specific antibody and specificity was satisfactory. The assay permits easy identification of determinant specificity on screening.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006510:Hepatitis B Antibodies / Q000096:biosynthesis / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011786:Quality Control; D013696:Temperature; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens