Abstract
We have expressed in bacteria a single-chain T cell receptor (scTCR) with specificity for an HIV gp120-derived peptide bound to the murine MHC-I molecule, H-2Dd. This scTCR consists of V alpha covalently linked to the VbetaCbeta domains that was solubilized, refolded, and purified in high yield. Specific binding of the scTCR to MHC/peptide complexes was demonstrated by surface plasmon resonance, with a Kd of 2 to 8 x 10(-6) M. This scTCR specifically inhibited T cell activation, and stained cell surface MHC/peptide complexes as measured by cytofluorimetry. The preservation of binding specificity by such a three-domain scTCR suggests that this structure is sufficient for specific MHC/peptide recognition and that this strategy will be of general use as applied to other TCR.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / chemistry
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Antibody Specificity
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Antigen-Antibody Reactions
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Base Sequence
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Circular Dichroism
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Epitopes / chemistry*
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Epitopes / immunology
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H-2 Antigens / chemistry*
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Hybridomas / immunology
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Immunoglobulins / chemistry
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Lymphocyte Activation
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Mice
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Molecular Sequence Data
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Peptides / chemistry*
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Peptides / immunology*
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Peptides / physiology
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Protein Binding / immunology
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Receptors, Antigen, T-Cell / biosynthesis
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Receptors, Antigen, T-Cell / chemistry*
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Receptors, Antigen, T-Cell / immunology
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Staining and Labeling
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T-Lymphocytes / immunology
Substances
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Antibodies, Monoclonal
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Epitopes
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H-2 Antigens
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Immunoglobulins
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Peptides
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Receptors, Antigen, T-Cell