Peptide deficient-MHC class I/chaperone compositions and methods
AssigneeThe Regents of the University of California
InventorNikolaos Sgourakis
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Abstract
Compositions that include stable peptide deficient MHC class I/chaperone complexes and methods of making and using such complexes are provided. In particular embodiments, such peptide deficient MHC class I/chaperone complexes are used to form peptide MHC class I (pMHC-I) multimers useful for high throughput applications, such as, for the detection of antigen specific T cells and characterization of T cell profiles in subjects.
Claims8
A method of making a library of peptide-MHC class I multimer complexes, each pMHC-I multimer complex in the library comprising an MHC class I multimer and a peptide of interest from a plurality of peptides of interest, the method comprising:
- a) incubating a plurality of MHC class I heavy chains, a plurality of MHC class I light chains, and a plurality of placeholder peptides under conditions wherein the plurality of MHC class I heavy chains, MHC class I light chains and placeholder peptides form a plurality of placeholder peptide-MHC class I (p*MHC-I) complexes;
- b) contacting the placeholder peptide-MHC class I (p*MHC-I) complexes with Tapasin Binding Protein Related (TAPBPR) chaperone to form peptide deficient-MHC class I/chaperone complexes;
- c) multimerizing the peptide deficient-MHC class I/chaperone complexes to form peptide deficient-MHC class I/chaperone multimer complex, wherein each peptide deficient-MHC class I/chaperone multimer complex comprises a backbone comprising a detectable label;
- and
- d) contacting each of the peptide deficient-MHC class I/chaperone multimer complexes with a peptide of interest from the plurality of peptides of interest, thereby forming a plurality of peptide-MHC class I multimer complexes.
Dependencies are inferred from the claim text.