A method for rapid screening of recombinant proteins for recognition by T lymphocytes.

Abstract

A simple, cost-effective method is described that allows rapid screening of recombinant protein sequences for their ability to stimulate T cells. Individual microcultures of E. coli each expressing a gene product or peptide sequence fused to protein A are grown in 96-well plates. Following lysis of the bacteria, the fusion peptide is readily captured with immobilized immunoglobulin in tissue culture wells. No further purification is required. T lymphocytes plus appropriate antigen-presenting cells are added directly to the wells and assayed for proliferation. The DNA in bacteria from wells stimulating T cell proliferation is then sequenced. The technique allows rapid mapping of T cell epitopes by facilitating screening of truncation mutants without extensive purification. Described here is a further application of the technique to study monosubstituted analogues of a known T cell epitope.

Cite this paper

@article{Hickling1992AMF, title={A method for rapid screening of recombinant proteins for recognition by T lymphocytes.}, author={Julian K Hickling and Kevin R. Jones and Bin Yuan and Jonathan B. Rothbard and Roland Buelow}, journal={European journal of immunology}, year={1992}, volume={22 8}, pages={1983-7} }