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

Ahmed S. Fahad, Cheng Yu Chung, Sheila N. Lopez Acevedo, Nicoleen Boyle, Bharat Madan, Matias F. Gutierrez-Gonzalez, Rodrigo Matus-Nicodemos, Amy D. Laflin, Rukmini R. Ladi, John Zhou, Jacy Wolfe, Sian Llewellyn-Lacey, Richard A. Koup, Daniel C. Douek, Henry H. Balfour, David A. Price, Brandon J. Dekosky

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


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.

Original languageEnglish (US)
Article numbergzab034
JournalProtein Engineering, Design and Selection
StatePublished - 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s) 2022. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail:


  • T cell receptor (TCR)
  • affinity-based screening
  • high-throughput TCRα:β sequencing
  • library immortalization

PubMed: MeSH publication types

  • Journal Article
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't


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