High Throughput pMHC-I Tetramer Library Production Using Chaperone Mediated Peptide Exchange
Sarah A Overall,
Jugmohit S Toor,
Alberto S Japp,
Michael R. Betts,
John M. Maris,
Posted 30 May 2019
bioRxiv DOI: 10.1101/653477 (published DOI: 10.1038/s41467-020-15710-1)
Posted 30 May 2019
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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