Peptide deficient-MHC class I/chaperone compositions and methods
AssigneeThe Regents of the University of California
InventorNikolaos G. 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.
Claims19
A method of a making peptide deficient-MHC class I/chaperone complex, the method comprising:
- a) incubating an MHC class I heavy chain, an MHC class I light chain and a placeholder peptide under conditions wherein the MHC class I heavy chain, the MHC class I light chain and the placeholder peptide form a placeholder peptide-MHC class I (p*MHC I) complex;
- and
- b) contacting the placeholder peptide-MHC class I complex with a dipeptide and Tapasin Binding Protein Related (TAPBPR) chaperone, thereby displacing the placeholder peptide from the placeholder peptide-MHC complex and forming the peptide deficient-MHC class I/chaperone complex.
A method of making a composition comprising a plurality of peptide-MHC class I (pMHC I) complexes, each complex comprising an MHC class I and a peptide 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) forming a plurality of peptide deficient-MHC class I/chaperone complexes by contacting the plurality of placeholder peptide-MHC class I complexes with a plurality of dipeptides and Tapasin Binding Protein Related (TAPBPR) chaperones;
- and
- c) contacting the plurality of peptide deficient-MHC class I/chaperone complexes with a plurality of peptides of interest, thereby forming the composition.
A method of making a composition comprising a plurality of peptide-MHC class I (pMHC-I) multimer complexes, each complex comprising an MHC class I multimer and a peptide 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 complexes (p*MHC-I);
- b) forming a plurality of peptide deficient MHC-chaperone complexes by contacting the plurality of placeholder peptide-MHC class I complexes (p*MHC-I) with a plurality of dipeptides and Tapasin Binding Protein Related (TAPBPR) chaperones;
- c) attaching the plurality of peptide deficient MHC class I/chaperone complexes to backbones, thereby forming a plurality of peptide deficient MHC class I/chaperone multimer complexes;
- d) contacting the plurality of peptide deficient-MHC class I/chaperone multimer complexes with a plurality of peptides of interest, thereby forming free chaperones and the plurality of peptide-MHC class I multimer complexes.
Dependencies are inferred from the claim text.