High throughput pMHC-I tetramer library production using chaperone-mediated peptide exchange

Sarah A. Overall, Jugmohit S. Toor, Stephanie Hao, Mark Yarmarkovich, Sara M. O’Rourke, Giora I. Morozov, Son Nguyen, Alberto Sada Japp, Nicolas Gonzalez, Danai Moschidi, Michael R. Betts, John M. Maris, Peter Smibert and Nikolaos G. Sgourakis ()
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Sarah A. Overall: University of California Santa Cruz
Jugmohit S. Toor: University of California Santa Cruz
Stephanie Hao: New York Genome Center
Mark Yarmarkovich: University of Pennsylvania
Sara M. O’Rourke: University of California Santa Cruz
Giora I. Morozov: University of California Santa Cruz
Son Nguyen: University of Pennsylvania
Alberto Sada Japp: University of Pennsylvania
Nicolas Gonzalez: University of California Santa Cruz
Danai Moschidi: University of California Santa Cruz
Michael R. Betts: University of Pennsylvania
John M. Maris: University of Pennsylvania
Peter Smibert: New York Genome Center
Nikolaos G. Sgourakis: University of California Santa Cruz

Nature Communications, 2020, vol. 11, issue 1, 1-13

Abstract: Abstract Peptide exchange technologies are essential for the generation of pMHC-multimer libraries used to probe diverse, polyclonal TCR repertoires in various settings. Here, using the molecular chaperone TAPBPR, we develop a robust method for the capture of stable, empty MHC-I molecules comprising murine H2 and human HLA alleles, which can be readily tetramerized and loaded with peptides of choice in a high-throughput manner. Alternatively, catalytic amounts of TAPBPR can be used to exchange placeholder peptides with high affinity peptides of interest. Using the same system, we describe high throughput assays to validate binding of multiple candidate peptides on empty MHC-I/TAPBPR complexes. Combined with tetramer-barcoding via a 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 antigen specificities in a single experiment. The new approach allows TCR/pMHC interactions to be interrogated easily at large scale.

Date: 2020
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-15710-1

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DOI: 10.1038/s41467-020-15710-1

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