Expression and secretion of T-cell receptor V alpha and V beta domains using Escherichia coli as a host

Abstract

An expression system for the production of recombinant T-cell receptor (TCR) variable domains would, inter alia, allow structural studies to be carried out and provide protein for the generation of anti-clonotypic antibodies. In this report the V alpha and V beta domain genes have been isolated from a T-cell hybridoma which is associated with the pathogenesis of experimental allergic encephalomyelitis (EAE) in the H-2u mouse. These have been expressed as secreted domains in Escherichia coli, using secretion vectors previously used for the production of immunoglobulin fragments. Both V alpha and V beta domains are secreted in milligram quantities into the culture supernatant, although the levels of the V alpha domain are about 10-20 fold higher than those of the V beta domain. This expression system offers a rapid route for the production of recombinant TCRs in soluble form.