Pipeline of TCR T-cell preclinical development. (A) Antigens can either be overexpressed self-antigens seen on normal tissues or those that belong to the cancer testis antigen family with restricted expression; neo-antigens arising from mutations and gene rearrangements; or those belonging to the minor histocompatibility complex. (B) Donor PBMCs containing either auto- or allo-HLA–restricted TCR-expressing T cells can be stimulated with antigenic peptides; and activated or reactive T cells can be identified, isolated, and expanded in the laboratory. (C) Sequencing of reactive cells identifies the precise αβ sequences of the TCR that are introduced into T cells after modifications to ensure proper pairing. Newly generated TCR T cells are tested for function and efficacy against tumor cells and tested for specificity and safety using in silico and experimental analyses. CDX, cell line-derived xenograft; DCs, dendritic cells; PDX, patient-derived xenograft. Figure created with BioRender.com: Gore S (2025); https://BioRender.com/o91f055