Immortalization and functional screening of natively paired human T cell receptor repertoires

Fahad, Ahmed S, Chung, Cheng-Yu, Lopez Acevedo, Sheila N, Boyle, Nicoleen, Madan, Bharat, Gutiérrez-González, Matias F, Matus-Nicodemos, Rodrigo, Laflin, Amy D, Ladi, Rukmini R, Zhou, John, Wolfe, Jacy, Llewellyn-Lacey, Sian, Koup, Richard A, Douek, Daniel C, Balfour Jr, Henry H, Price, David A ORCID: https://orcid.org/0000-0001-9416-2737 and DeKosky, Brandon J 2022. Immortalization and functional screening of natively paired human T cell receptor repertoires. Protein Engineering, Design & Selection 35 , gzab034. 10.1093/protein/gzab034

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Abstract

Functional analyses of the T cell receptor (TCR) landscape can reveal critical information about protection from disease and molecular responses to vaccines. However, it has proven difficult to combine advanced next-generation sequencing technologies with methods to decode the peptide-major histocompatibility complex (pMHC) specificity of individual TCRs. We developed a new high-throughput approach to enable repertoire-scale functional evaluations of natively paired TCRs. In particular, we leveraged the immortalized nature of physically linked TCRα:β amplicon libraries to analyze binding against multiple recombinant pMHCs on a repertoire scale, and to exemplify the utility of this approach, we also performed affinity-based functional mapping in conjunction with quantitative next-generation sequencing to track antigen-specific TCRs. These data successfully validated a new immortalization and screening platform to facilitate detailed molecular analyses of disease-relevant antigen interactions with human TCRs.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Medicine
Publisher:	Oxford University Press
ISSN:	1741-0126
Date of First Compliant Deposit:	17 February 2023
Date of Acceptance:	29 December 2021
Last Modified:	12 May 2023 21:41
URI:	https://orca.cardiff.ac.uk/id/eprint/157135

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