Requirements for screening recombinant DNA libraries for T cell epitope expression.
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.
Guttinger
M
M
Central Research Units, F. Hoffmann-La Roche & Co. Ltd., Basel, Switzerland.
Certa
U
U
Pink
J R
JR
Sinigaglia
F
F
D002999:Clone Cells; D004247:DNA / Q000032:analysis*; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D013601:T-Lymphocytes / Q000276:immunology*
D000939:Epitopes; D004247:DNA