Immortalization and functional screening of natively paired human T cell receptor repertoires. (17th February 2022)

• Record Type: Journal Article
• Authors: 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; DeKosky, Brandon J
• Is Part Of: Protein engineering, design & selection. Volume 35(2022)
• Publication Date: 2022-02-17
• Subjects: T cell receptor (TCR) -- high-throughput TCRα:β sequencing -- library immortalization -- affinity-based screening; Protein engineering -- Periodicals; 660.63
• DOI: 10.1093/protein/gzab034
• Languages: English
• ISSNs: 1741-0126

Abstract

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.