High-throughput screening of functional neo-antigens and their specific TCRs via the Jurkat reporter system combine with droplet microfluidics

T cell receptor-engineered T cells (TCR-T) therapy would be one of the most promising technologies in solid tumor immune therapy benefit from recognizing tumor neo-antigens both in cell surface and intracellular. However, time-consuming steps and low accuracy to screen functional antigens and their specific TCRs from thousands of candidate libraries may severely limits their practical application. Here, we developed an integrated platform which combine with Jurkat reporter system, DNA barcoding and droplet microfluidics to enable the library-on-library functional antigens and TCR screening. Next, analysis of T cell activation related genes and DNA barcode based on single-cell RNA-sequencing can determine the specific recognition between the antigens and their functional TCRs. Our proof-of principle study demonstrates that screening antigens and their functional TCRs simultaneously with high sensitivity and specificity in unbiased library-on-library screening can be achieved, which could significantly reduce the time required in operation, and accelerate progress toward cancer treatment. Moreover, it also be expected to expand the research filed of screening the cross-reactivity and off-target recognition of candidate antigens and TCRs in libraries before clinical application. Keywords: High throughput screening, functional TCR screening, jurkat reporter, droplet-based microfluidics, single-cell transcriptome ### Competing Interest Statement The authors have declared no competing interest.