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US 10,481,158 B2Granted
Compositions and methods for screening T cells with antigens for specific populations
AssigneeCalifornia Institute of Technology
InventorsJames R. Heath, Songming Peng
Life of this patent
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Abstract
Compositions and methods for isolating patient-derived antigen-specific T cells include an antigen complex having a polynucleotide barcoded nanoparticle sorting agent complexed with a peptide-loaded streptavidin major histocompatability complex (MHC) tetramer, the barcoding technology allowing for high fidelity screening of a library of the antigen complexes to readily isolate and identify antigen-specific T cells.
Claims19
An antigen complex, comprising:
- a nanoparticle sorting agent comprising:
- a nanoparticle;
- a polynucleotide detection tag having at least one coding region, the polynucleotide detection tag being conjugated to the nanoparticle;
- and
- a first polynucleotide hybridization domain conjugated to the polynucleotide detection tag;
- and
- a peptide-loaded streptavidin major histocompatability complex (MHC) tetramer, comprising:
- a modified streptavidin protein;
- four biotin-modified MHC proteins each independently conjugated to the modified streptavidin protein;
- an antigen peptide bound to the biotin-modified MHC proteins;
- and
- a second polynucleotide hybridization domain different from the first polynucleotide hybridization domain and conjugated to the modified streptavidin protein,
- the nanoparticle sorting agent being linked to the peptide-loaded streptavidin MHC tetramer by hybridization of the first polynucleotide hybridization domain to the second polynucleotide hybridization domain.
10 more elements
A kit for detecting neoantigen-specific T cells, comprising:
- an antigen complex, comprising:
- a nanoparticle sorting agent comprising:
- a nanoparticle;
- a polynucleotide detection tag having at least one coding region, the polynucleotide detection tag being conjugated to the nanoparticle;
- and
- a first polynucleotide hybridization domain conjugated to the polynucleotide detection tag;
- and
- a peptide-loaded streptavidin major histocompatability complex (MHC) tetramer, comprising:
- a modified streptavidin protein;
- four biotin-modified MHC proteins each independently conjugated to the modified streptavidin protein;
- an antigen peptide bound to the biotin-modified MHC proteins;
- and
- a second polynucleotide hybridization domain different from the first polynucleotide hybridization domain and conjugated to the modified streptavidin protein,
- the nanoparticle sorting agent being linked to the peptide-loaded streptavidin MEW tetramer by hybridization of the first polynucleotide hybridization domain to the second polynucleotide hybridization domain;
- and
- a decoding polynucleotide that is capable of hybridizing to the at least one coding region of the polynucleotide detection tag.
13 more elements
A method for isolating neo-antigen-specific T cells for a tumor in a subject, comprising:
- identifying candidate T cell epitopes for the tumor using a major histocompatiblity complex (MHC) binding algorithm;
- synthesizing antigen peptides corresponding to the candidate T cell epitopes;
- preparing a library of antigen complexes using the antigen peptides, the antigen complexes each comprising:
- a nanoparticle sorting agent comprising:
- a nanoparticle;
- a polynucleotide detection tag having at least one coding region, the polynucleotide detection tag being conjugated to the nanoparticle;
- and
- a first polynucleotide hybridization domain conjugated to the polynucleotide detection tag;
- and
- a peptide-loaded streptavidin major histocompatability complex (MHC) tetramer, comprising:
- a modified streptavidin protein;
- four biotin-modified MHC proteins each independently conjugated to the modified streptavidin protein;
- the antigen peptide bound to the biotin-modified MHC proteins;
- and
- a second polynucleotide hybridization domain different from the first polynucleotide hybridization domain and conjugated to the modified streptavidin protein,
- the nanoparticle sorting agent being linked to the peptide-loaded streptavidin MHC tetramer by hybridization of the first polynucleotide hybridization domain to the second polynucleotide hybridization domain;
- each of the antigen complexes having a different antigen peptide and different polynucleotide detection tag than any other of the antigen complexes in the plurality of antigen complexes;
- incubating the library of antigen complexes with TILs or PBMCs from the subject;
- and
- separating paired T cells from unpaired T cells, the paired T cells comprising those T cells that have paired with any of the antigen peptides of any of the antigen complexes in the library of antigen complexes.
17 more elements
Dependencies are inferred from the claim text.