Peptide-receptive MHC-I complex compositions and methods
AssigneeThe Regents of the University of California
InventorNikolaos G. Sgourakis
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Abstract
Compositions that include peptide receptive MHC class I complexes and methods of making and using such complexes are provided. In particular embodiments, such peptide receptive MHC class I 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.
Claims23
A method of making a plurality of peptide receptive MHC-I complexes each of which can accept a peptide of interest, each peptide receptive MHC-I complex comprising an MHC class I heavy chain and a β2-microglobulin, the method comprising:
- a) incubating a plurality of MHC class I heavy chains;
- a plurality of β2-microglobulins;
- and a plurality of placeholder peptides, under conditions wherein the plurality of MHC class I heavy chains, the β2-microglobulins and the plurality placeholder peptides form a plurality of placeholder peptide-MHC class I (p*MHC-I) complexes, wherein all of the MHC class I heavy chains in the plurality of MHC class I heavy chains are HLA-A*24:02 and all of the placeholder peptides in the plurality of placeholder peptides consist of the amino acid sequence YPLFGWCF (SEQ ID NO:1), or wherein all of the MHC class I heavy chains in the plurality of MHC class I heavy chains are HLA-A*68:02 and all of the placeholder peptides in the plurality of placeholder peptides consist of the amino acid sequence LFGPVYV (SEQ ID NO:2);
- and
- b) contacting the p*MHC-I complexes with a plurality of Tapasin Binding Protein Related (TAPBPR) chaperones, thereby creating the plurality of peptide receptive MHC-I complexes.
A method of making a plurality of peptide-MHC class I (pMHC-I) complexes, each complex comprising an MHC Class I heavy chain, a β2-microglobulin, and a peptide of interest, the method comprising:
- a) incubating a plurality of MHC class I heavy;
- a plurality of β2-microglobulins;
- and a plurality of placeholder peptides, under conditions wherein the plurality of MHC class I heavy chains, the plurality β2-microglobulins and the plurality placeholder peptides form a plurality of placeholder peptide-MHC class I (p*MHC-I) complexes, wherein all of the MHC class I heavy chains in the plurality of MHC class I heavy chains are HLA-A*24:02 and all of the placeholder peptides in the plurality of placeholder peptides consist of the amino acid sequence YPLFGWCF (SEQ ID NO:1), or wherein all of the MHC class I heavy chains in the plurality of MHC class I heavy chains are HLA-A*68:02 and all of the placeholder peptides in the plurality of placeholder peptides consist of the amino acid sequence LFGPVYV (SEQ ID NO:2);
- b) forming a plurality of peptide receptive MHC-I complexes by contacting the plurality of p*MHC-I complexes with a plurality of TAPBPR chaperones;
- and
- c) contacting the plurality of peptide receptive MHC-I complexes with a plurality of peptides of interest, thereby forming the plurality of pMHC-I complexes.
A method of making a plurality of peptide-MHC class I (pMHC-I) multimers, each pMHC-I multimer comprising a plurality of pMHC-I complexes, the method comprising:
- a) incubating a plurality of MHC class I heavy;
- a plurality of β2-microglobulins;
- and a plurality of placeholder peptides, under conditions wherein the plurality of MHC class I heavy chains, the β2-microglobulins and the plurality of placeholder peptides form a plurality of placeholder peptide-MHC class I (p*MHC-I) complexes, wherein all of the MHC class I heavy chains in the plurality of MHC class I heavy chains are HLA-A*24:02 and all of the placeholder peptides in the plurality of placeholder peptides consist of the amino acid sequence YPLFGWCF (SEQ ID NO:1), or wherein all of the MHC class I heavy chains in the plurality of MHC class I heavy chains are HLA-A*68:02 and all of the placeholder peptides in the plurality of placeholder peptides consist of the amino acid sequence LFGPVYV (SEQ ID NO:2);
- b) contacting the plurality of p*MHC-I complexes with a plurality TAPBPR chaperones, thereby forming a plurality of peptide receptive MHC-I complexes;
- c) attaching the plurality of peptide receptive MHC-I complexes to multimer backbones, thereby forming a plurality of peptide receptive MHC-I multimers;
- and
- d) contacting the plurality of peptide receptive MHC-I multimers with a plurality of peptides of interest, thereby forming a plurality of pMHC-I multimers.
Dependencies are inferred from the claim text.