A NOVEL AND EFFICIENT ROUTE FOR THE ISOLATION OF ANTIBODIES THAT RECOGNIZE T-CELL RECEPTOR

Studies of the T cell repertoire have been hindered by the lack of ant ibodies that recognise V region families, particularly for V alpha reg ions. In this report, single chain Fv (scFv) fragments have been isola ted that recognise both recombinant V alpha s and native V alpha s on the surface of T cells. Mice have been immunised with purified soluble T cell receptors (TCRs) and antibody heavy and light chain variable d omain (VH and VL, respectively) genes isolated from splenocytes using the polymerase chain reaction (PCR). The VH and VL genes have been ass embled as scFv gene libraries and a bacteriophage display system used to isolate scFvs that recognise a soluble V alpha. Five scFvs have bee n purified and characterised in detail using enzyme-linked immunosorbe nt assays (ELISAs) and flow cytometry. Three of these five scFvs recog nise native V alpha s on the surface of T cell hybridomas. This method therefore offers a rapid route to the generation of scFvs that recogn ise native TCRs and can readily be extended to the production of anti- human TCR antibodies for use in therapy and diagnosis. Copyright (C) 1 996 Elsevier Science Ltd.