A solid-phase immunofluorescence assay (SIFA) for screening antigen-specific hybridomas.
A solid-phase immunofluorescence assay (SIFA) is described for screening of monoclonal hybridoma antibodies against soluble antigens. The test is simple, needs a small amount of material (20-100 ng of labeled antigen per sample) and allows a large number of samples (500-1 000) to be tested in 2 days. The assay is performed in microtitration plates with small cellulose nitrate discs as the solid phase. Anti-mouse IgG, the sample to be tested and FITC-labeled antigen are successively incubated with the discs. The fluorescence intensity of the surface of the discs is then measured with a microfluorometer. The assay has been used to select hybridomas against human serum albumin. Comparison of the immunofluorescence assay with a radioimmunoassay using conventional antisera and hybridoma antibodies similar sensitivity for both assays.
D000818:Animals; D000939:Epitopes; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007106:Immune Sera / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D011863:Radioimmunoassay; D012709:Serum Albumin / Q000276:immunology*
D000939:Epitopes; D007106:Immune Sera; D012709:Serum Albumin