Characterization of a Single-chain Antibody to the 
-Chain of the T Cell Receptor (*)
- § To whom correspondence should be addressed: Dept. of Biochemistry, University of Illinois, 600 S. Matthews Ave., Urbana, IL 61801-3792. Tel.: 217-244-2821; Fax: 217-244-5858.
Abstract
In this report the V
and V
genes of the anti-T cell receptor (TCR) antibody KJ16, which recognizes the TCR Vβ8.1 and Vβ8.2 regions in mice, were cloned
and expressed as a single-chain antibody (scFv) in Escherichia coli. A 29-kDa protein was obtained after renaturation from inclusion bodies. The KJ16 scFv had a relative affinity for the native
TCR that was slightly higher than KJ16 Fab fragments. The scFv and Fab fragments of the KJ16 antibody, together with monovalent
forms of two other anti-TCR antibodies, were evaluated as antagonists of the T cell-mediated recognition of a peptide-class
I complex or of a superantigen, Staphylococcus enterotoxin B (SEB) bound to a class II product. Each of the anti-TCR antibodies was efficient at inhibiting the recognition
of the SEB-class II complex. In contrast, only the clonotypic antibody, which binds to epitopes on both the Vβ and Vα regions,
inhibited the recognition of peptide-class I complex. We conclude that the TCR binding site for the SEB-class II ligand encompasses
a larger surface area than the TCR binding site for the peptide-class I ligand.
Footnotes
-
↵* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. This work was supported by National Institutes of Health Grant AI35990.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) U34924[GenBank].
-
↵1 The abbreviations used are:
- TCR
-
T cell receptor
- scFv
-
single-chain antibody binding domain
- scTCR
-
single-chain T cell receptor
- CTL
-
cytotoxic T lymphocyte
- CDR
-
complementarity-determining region
- MHC
-
major histocompatibility complex
- Fab
-
antigen binding fragment derived from papain digestion of Ig molecule
- Fc
-
effector region of an antibody derived from papain digestion of Ig molecule
- V
and V
-
variable regions of Ig heavy and light chains, respectively
- Vα and Vβ
-
variable regions of the TCR α and β chains, respectively
- IC
-
the concentration required to inhibit 50% of the binding to 2C TCR
- SEB
-
Staphylococcus enterotoxin B
- FITC
-
5-aminofluorescein isothiocyanate
- FITC/Fab
-
Fab fragments labeled with fluorescein
- PAGE
-
polyacrylamide gel electrophoresis
- ELISA
-
enzyme-linked immunosorbant assay
- HPLC
-
high pressure liquid chromatography
- PBS
-
phosphate-buffered saline.
-
↵2D. M. Kranz, unpublished data.
-
↵3B. K. Cho and D. M. Kranz, unpublished data.
-
- Received July 24, 1995.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
