Reactive T Cell Receptor Identification Before Expansion

The patent applies preliminary action by identifying and isolating tumor-reactive T cells and their TCRs before the cell expansion process. Gene expression profiling and TCR sequencing are performed on pre-enriched T cells to capture the reactive clones early, avoiding the exhaustion that occurs during prolonged in vitro expansion. This allows subsequent production of T cell products based on the identified TCRs without compromising the reactive clones themselves.

The patent applies copying by creating T cell products that express TCRs identified from the patient's own T cells, rather than directly expanding the original reactive clones. The TCR sequences are determined through sequencing, and these sequences are then used to engineer T cells (either autologous or allogeneic) that carry the identified reactive TCRs. This copying approach allows mass production of uniform, specific T cell products without the clone loss that occurs during extensive in vitro expansion of primary T cells.

The patent applies universality by using gene expression profiling (such as scRNA-seq) as a multi-functional tool that simultaneously achieves multiple objectives: identifying reactive T cells, determining their TCR repertoires, characterizing their activation state, and discovering biomarkers of reactivity. This single approach provides comprehensive information that would otherwise require multiple separate assays, thereby managing complexity while maintaining high identification precision.

The patent applies merging by combining TCR sequencing with gene expression profiling in a unified analytical workflow. Both TCR alpha and beta chain sequences are determined simultaneously with gene expression data from the same T cell population. This integrated approach allows parallel processing of multiple parameters from the same samples, reducing the total time and resource requirements compared to performing separate sequencing and expression studies.