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Requirements for screening recombinant DNA libraries for T cell epitope expression.

M Guttinger·U Certa·J R Pink·F Sinigaglia
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Abstract

The possibility of screening cDNA expression libraries with T cell clones was investigated. The model system was based on human T cell clones specific for the recombinant malaria protein 190L, which was expressed fused to beta-galactosidase in lambda gt11. Several membranes were tested for their capacity to bind antigen and stimulate T cell proliferation. Pretreatment of membranes with DMSO and/or sonication to release the antigen improved the sensitivity of the assay. Under optimal conditions, T cell proliferation in response to antigen bound to a low protein binder membrane was comparable to that observed with the antigen in solution. A dot-blot type apparatus was designed for screening large numbers of plaques with T cells. The technical problems of this approach, its requirements and possible applications are discussed.

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Clone CellsDNAEpitopesHumansT-LymphocytesJournal Article

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M Guttinger, U Certa, J R Pink, F Sinigaglia. 1989-07-26. Requirements for screening recombinant DNA libraries for T cell epitope expression.. https://doi.org/10.1016/0022-1759(89)90164-6

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