Affinity and functional based screening of t cell receptor repertoire using three-dimensional shaped particles

Abstract

The ability to selectively bind to antigenic peptides and secrete cytokines and other biomolecules can define T cells with therapeutic potential in T cell receptor (TCR) immunotherapies. Hydrogel microparticles (nanovials) having cavities for holding T cells are coated with peptide-major histocompatibility complex (pMHC) monomers to isolate antigen-reactive T cells. T cells are captured and activated by pMHCs inducing the secretion of effector molecules that are captured on nanovials that permits sorting based on both binding and function. The TCRs of sorted cells on nanovials are sequenced, recovering paired αβ-chains using single-cell sequencing. By labeling nanovials having different pMHCs with unique oligonucleotide-barcodes and secretions with oligo-barcoded detection antibodies one can accurately link TCR sequences to specific targets and rank each TCR based on the corresponding T cell's secretion level. An expanded repertoire of functional TCRs targeting viral antigens were discovered along with rare TCRs with activity against prostate cancer-specific antigens.

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