Characterization of a single-chain antibody to the beta-chain of the T cell receptor.
In this report the VH and VL genes of the anti-T cell receptor (TCR) antibody KJ16, which recognizes the TCR V beta 8.1 and V beta 8.2 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 V beta and V alpha 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.
Cho
B K
BK
Department of Biochemistry, University of Illinois, Urbana 61801-3792, USA.
Schodin
B A
BA
Kranz
D M
DM
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D003001:Cloning, Molecular; D003602:Cytotoxicity, Immunologic; D004768:Enterotoxins / Q000276:immunology; D006649:Histocompatibility Antigens / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D051379:Mice; D015335:Molecular Probes; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D017422:Sequence Analysis, DNA; D018089:Superantigens / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D004768:Enterotoxins; D006649:Histocompatibility Antigens; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D015335:Molecular Probes; D010455:Peptides; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; D018089:Superantigens; C031818:enterotoxin B, staphylococcal