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High Throughput pMHC-I Tetramer Library Production Using Chaperone Mediated Peptide Exchange

By Sarah A Overall, Jugmohit S Toor, Stephanie Hao, Mark Yarmarkovich, Son Nguyen, Alberto S Japp, Danai Moschidi, Michael R Betts, John M. Maris, Peter Smibert, Nikolaos G Sgourakis

Posted 30 May 2019
bioRxiv DOI: 10.1101/653477

Peptide exchange technologies are essential for the generation of pMHC-multimer libraries, used to probe highly diverse, polyclonal TCR repertoires. Using the molecular chaperone TAPBPR, we present a robust method for the capture of stable, empty MHC-I molecules which can be readily tetramerized and loaded with peptides of choice in a high-throughput manner. Combined with tetramer barcoding using multi-modal cellular indexing technology (ECCITE-seq), our approach allows a combined analysis of TCR repertoires and other T-cell transcription profiles together with their cognate pMHC-I specificities in a single experiment.

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