Use of a solid-phase 3H-radioimmunoassay for the measurement of immunoglobulin produced in short-term cultures of antibody-secreting cells.
A slid-phase radioimmunoassay (RIA) for assaying immunoglobulin produced from antibody-secreting myeloma, hybridoma and immune spleen cells is described. Specific antibody is detected by culturing antibody-producing cells on antigen-coated flexible polyvinylchloride microtiter wells, washing away the cells, and measuring the bound specific antibody with tritiated affinity-purified anti-isotype reagents. Antibody produced from 10(3) myeloma cells can be detected with as little as 4 h of incubation. With 24-48 h of incubation, antibody from as few as approximately 3-15 myeloma, hybridoma or immune spleen plaque-forming cells (PFC) can be detected. This culture-well RIA has certain distinct advantages over the hemolytic PFC assay and RIA assays in which antibody in culture supernatants is measured.
D000818:Animals; D000918:Antibody Specificity; D000921:Antibody-Producing Cells / Q000032:analysis*; D000941:Antigens; D001666:Binding Sites, Antibody; D002417:Cattle; D002478:Cells, Cultured; D006825:Hybridomas / Q000032:analysis / Q000276:immunology; D007070:Immunoglobulin A / Q000032:analysis / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000032:analysis; D007136:Immunoglobulins / Q000032:analysis* / Q000096:biosynthesis; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D010954:Plasmacytoma / Q000032:analysis / Q000276:immunology; D011863:Radioimmunoassay; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014301:Trinitrobenzenes / Q000276:immunology
D000941:Antigens; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007136:Immunoglobulins; D014301:Trinitrobenzenes; D012710:Serum Albumin, Bovine