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A solid-phase immunofluorescence assay (SIFA) for screening antigen-specific hybridomas.

B Micheel·U Karsten·H Fiebach
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Abstract

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.

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AnimalsEpitopesFemaleFluorescent Antibody TechniqueHumansHybridomasImmune SeraMiceMice, Inbred BALB CRadioimmunoassaySerum AlbuminJournal Article

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B Micheel, U Karsten, H Fiebach. 1981. A solid-phase immunofluorescence assay (SIFA) for screening antigen-specific hybridomas.. https://doi.org/10.1016/0022-1759(81)90330-6

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