Method for identifying cytotoxic T-cell epitopes
OwnerCharles A. Nicoletteinventor
Life of this patent
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Abstract
A method for isolating bio-active molecules from complex, rationally designed oligopeptide libraries which elicit cytolytic activity from cloned cytotoxic T lymphocytes (CTLs). The method allows the simultaneous screening of multiple CTL lines against peptide libraries synthesized on solid phase support. The method to identify CTL-reactive oligopeptides yields products of therapeutic value such as vaccines in treating cancers, viral diseases, and autoimmune diseases, as well as to identify useful clinical diagnostic reagents with a reduction in assay time and increase in throughput.
Claims28
A method for identifying a cytotoxic T cell epitope comprising the steps in order of:
- a) contacting a population of at least two cytotoxic T cells having the same MHC-haplotype restriction with
- i) a library of molecules attached to solid phase supports by a releasable linker, wherein each solid phase support is attached to a single species of molecule, and wherein the structure of the molecule can be determined, which library of molecules contains a conserved structural motif corresponding to a structural motif characteristic of peptides that associate with the MHC-haplotype to which the cytotoxic T cells are restricted;
- and
- ii) antigen presentation means, which antigen presentation means correspond to the MHC-haplotype to which the cytotoxic T cells are restricted;
- wherein the solid phase supports of the library are in separate fractions;
- b) cleaving at least a portion of the releasable linker so as to release at least a portion of the molecule;
- c) evaluating whether the cytotoxic T cells recognize a molecule present in one or more of the fractions of the library of molecules;
- d) isolating one or more solid phase support(s) from the fractions;
- and
- e) determining the structure of a molecule on a solid phase support isolated from the fraction.
Dependencies are inferred from the claim text.