IDENTIFICATION OF ANTIGEN EPITOPES AND IMMUNE SEQUENCES RECOGNIZING THE ANTIGENS

摘要

Methods, compositions, and reaction mixtures are provided for identifying a T cell receptor (TCR) and an epitope peptide that specifically binds the TCR. Methods, compositions, and reaction mixtures are also provided for identifying a plurality of T cell receptors and corresponding epitope peptides that specifically bind the T cell receptors. In some cases, the plurality of T cell receptors and corresponding epitope peptides can be identified in a highly parallel manner.

主权项

1. A method for identifying a T cell receptor (TCR) and an epitope peptide that specifically binds the TCR, the method comprising:
a) providing a plurality of mixture partitions comprising a population of T cells, the T cells comprising a plurality of different TCRs having a TCR heavy chain encoded by a TCR heavy chain nucleic acid and a TCR light chain encoded by a TCR light chain nucleic acid, wherein individual mixture partitions of the plurality of mixture partitions comprise:
i) a portion of the population of T cells, wherein the portion of T cells comprises one or more TCRs that specifically bind to a portion of a plurality of epitope peptides present in the mixture partition;ii) a plurality of partition-specific barcodes; andiii) a plurality of epitope nucleic acids encoding the epitope peptides; b) in the plurality of mixture partitions, generating barcoded nucleic acids by attaching the partition-specific barcodes to the TCR heavy chain, TCR light chain, and epitope nucleic acids; c) pooling the barcoded nucleic acids to form a pooled mixture; and d) sequencing the barcoded nucleic acids in the pooled mixture and thereby identifying the sequence of the heavy and light chain of the TCR that specifically binds the epitope peptide and the sequence of the epitope peptide that specifically binds the TCR by identifying heavy and light chain nucleic acids and epitope nucleic acids having matching partition-specific barcodes.