Abstract

In this webinar, we will discuss T-cell receptor (TCR) mapping of antigenic peptides (TCR-MAP), a cell-based method to discover T-cell antigens using a synthetic TCR-stimulated circuit in immortalized T cells. TCRs with unknown specificity are queried against a large, barcoded peptide library displayed on antigen-presenting cells (APCs). Upon TCR recognition of the peptide–MHC complex presented by APCs, sortaseA is induced and activated, which enables covalent biotinylation of the adjacent APCs, allowing for their downstream sorting. These live, tagged APCs can then be directly purified for deconvolution by sequencing.

TCR-MAP is a high-throughput method that can accurately capture unbiased reactivities against the complete human, mouse, or viral proteome or any genetically encoded peptide library of choice. Moreover, it is highly sensitive and can reproducibly discover both high-affinity and low-affinity TCR antigens. Application of this technology has the potential to enhance T-cell antigen discovery efforts in the context of cancer, infectious diseases, and autoimmunity.

Highlights:

• High-throughput T-cell antigen discovery technology platform.

• Recognition-induced tagging of antigen-presenting cells.

• Human T-cell receptors with unknown specificity queried against barcoded peptide libraries.