PMID: 7009749Jan 1, 1980Paper

Screening and replica plating of anti-hapten hybridomas with a transfer template hemolytic spot assay

Journal of Immunological Methods
R B BankertL Powers

Abstract

A localized hemolysis in gel assay is described for screening microcultures of hybridomas for the production of anti-hapten antibody. The keys to the rapid screening assay reported here are a special fenestrated transfer template and an improved hapten conjugated target cell. The transfer template is a 96-well plate with a calibrated hole in the bottom center of each well. To assay for anti-hapten antibodies, the transfer template is positioned over a 96-well microculture plate containing the growing hybridomas. After making contact with the tissue culture supernatant each orifice of the transfer template retains approximately 2 microliter of tissue culture supernatant. The transfer template is then placed onto an assay slide containing a thin layer of hapten conjugated target erythrocytes incorporated into agarose. After incubation with an anti-immunoglobulin and complement, areas of localized hemolysis in the gel indicate hybridomas which are secreting anti-hapten antibodies. The assay detects as little as 10 pg of antibody. Since the transfer template can be used as a replica plate, one can repeatedly transfer samples to various slides which contain either different hapten target cells or different hapten analog inhibitors i...Continue Reading

Citations

Oct 1, 1982·Developmental Biology·B L Hylander, R G Summers
Jan 1, 1982·Journal of Immunological Methods·M L CohnM B Rittenberg
Apr 1, 1985·Molecular Immunology·A L BarsoumE A Davidson
Apr 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·Y H Jou, R B Bankert
Jun 1, 1982·Cancer Treatment Reviews·C S Foster

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