We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
High throughput pMHC-I tetramer library production using chaperone-mediated peptide exchange.
- Authors
Overall, Sarah A.; Toor, Jugmohit S.; Hao, Stephanie; Yarmarkovich, Mark; Sara M. O'Rourke; Morozov, Giora I.; Nguyen, Son; Japp, Alberto Sada; Gonzalez, Nicolas; Moschidi, Danai; Betts, Michael R.; Maris, John M.; Smibert, Peter; Sgourakis, Nikolaos G.
- 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. Peptide-MHC (pMHC) tetramers are important tools for probing T cell repertoire and adaptive immune responses. Here the authors use a molecular chaperone, TAPBPR, to develop a high-throughput, multiplexible platform for pMHC tetramer generation to facilitate simultaneous assessments of T cell repertoire/antigen specificity and transcriptome.
- Subjects
BINDING site assay; PEPTIDES; EXCHANGE; T cells; MOLECULAR chaperones; IMMUNE response
- Publication
Nature Communications, 2020, Vol 11, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-020-15710-1