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A novel and efficient route for the isolation of antibodies that recognise T cell receptor Vαs

A novel and efficient route for the isolation of antibodies that recognise T cell receptor Vαs
A novel and efficient route for the isolation of antibodies that recognise T cell receptor Vαs

Studies of the T cell repertoire have been hindered by the lack of antibodies that recognise V region families, particularly for Vα regions. In this report, single chain Fv (scFv) fragments have been isolated that recognise both recombinant Vαs and native Vαs on the surface of T cells. Mice have been immunised with purified soluble T cell receptors (TCRs) and antibody heavy and light chain variable domain (VH and VL, respectively) genes isolated from splnocytes using the polymerase chain reaction (PCR). The VH and VL genes have been assembled as scFv gene libraries and a bacteriophage display system used to isolate scFvs that recognise a soluble Vα. Five scFvs have been purified and characterised in detail using enzyme-linked immunosorbent assays (ELISAs) and flow cytometry. Three of these five scFvs recognise native Vαs on the surface of T cell hybridomas. This method therefore offers a rapid route to the generation of scFvs that recognise native TCRs and can readily be extended to the production of anti-human TCR antibodies for use in therapy and diagnosis.

Antibodies, Phage display, Single chain Fv, T cell receptor
0161-5890
493-502
Popov, Sergei
15a37b99-253a-43a4-b825-19cd4e9b150e
Hubbard, J. Gregory
8394863f-e859-4e83-8d13-7f19e5760264
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Popov, Sergei
15a37b99-253a-43a4-b825-19cd4e9b150e
Hubbard, J. Gregory
8394863f-e859-4e83-8d13-7f19e5760264
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc

Popov, Sergei, Hubbard, J. Gregory and Ward, E. Sally (1996) A novel and efficient route for the isolation of antibodies that recognise T cell receptor Vαs. Molecular Immunology, 33 (6), 493-502. (doi:10.1016/0161-5890(96)00020-X).

Record type: Article

Abstract

Studies of the T cell repertoire have been hindered by the lack of antibodies that recognise V region families, particularly for Vα regions. In this report, single chain Fv (scFv) fragments have been isolated that recognise both recombinant Vαs and native Vαs on the surface of T cells. Mice have been immunised with purified soluble T cell receptors (TCRs) and antibody heavy and light chain variable domain (VH and VL, respectively) genes isolated from splnocytes using the polymerase chain reaction (PCR). The VH and VL genes have been assembled as scFv gene libraries and a bacteriophage display system used to isolate scFvs that recognise a soluble Vα. Five scFvs have been purified and characterised in detail using enzyme-linked immunosorbent assays (ELISAs) and flow cytometry. Three of these five scFvs recognise native Vαs on the surface of T cell hybridomas. This method therefore offers a rapid route to the generation of scFvs that recognise native TCRs and can readily be extended to the production of anti-human TCR antibodies for use in therapy and diagnosis.

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More information

Accepted/In Press date: 13 February 1996
Published date: April 1996
Keywords: Antibodies, Phage display, Single chain Fv, T cell receptor

Identifiers

Local EPrints ID: 425113
URI: http://eprints.soton.ac.uk/id/eprint/425113
ISSN: 0161-5890
PURE UUID: acb0fb9f-0a5f-442c-a27c-346d8977fd69
ORCID for E. Sally Ward: ORCID iD orcid.org/0000-0003-3232-7238

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Date deposited: 11 Oct 2018 16:30
Last modified: 18 Mar 2024 03:48

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Contributors

Author: Sergei Popov
Author: J. Gregory Hubbard
Author: E. Sally Ward ORCID iD

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