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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