Immortalization and Functional Screening of Natively Paired Human T Cell Receptor Repertoires

Fahad, A. S.; Yu Chung, C.; Lopez Acevedo, S. N.; Boyle, N.; Madan, B.; Gutierrez-Gonzalez, M. F.; Matus-Nicodemos, R.; Laflin, A. D.; Ladi, R. R.; Zhou, J.; Wolfe, J.; Llewellyn-Lacey, S.; Doueck, D.; Balfour, H. H.; Price, D. A.; DeKosky, B. J.

doi:10.1101/2021.10.19.465057

bioRxiv · 10.1101/2021.10.19.465057

Immortalization and Functional Screening of Natively Paired Human T Cell Receptor Repertoires

Fahad, A. S.·Yu Chung, C.·Lopez Acevedo, S. N.·Boyle, N.·Madan, B.·Gutierrez-Gonzalez, M. F.·Matus-Nicodemos, R.·Laflin, A. D.·Ladi, R. R.·Zhou, J.·Wolfe, J.·Llewellyn-Lacey, S.·Doueck, D.·Balfour, H. H.·Price, D. A.·DeKosky, B. J.

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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. Here 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 TCR:{beta} amplicon libraries to analyze binding against multiple recombinant pMHCs on a repertoire scale. 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 human TCRs against diverse antigen targets associated with health, vaccination, or disease.

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Fahad, A. S., Yu Chung, C., Lopez Acevedo, S. N., Boyle, N., Madan, B., Gutierrez-Gonzalez, M. F., Matus-Nicodemos, R., Laflin, A. D., Ladi, R. R., Zhou, J., Wolfe, J., Llewellyn-Lacey, S., Doueck, D., Balfour, H. H., Price, D. A., DeKosky, B. J.. 2021-10-21. Immortalization and Functional Screening of Natively Paired Human T Cell Receptor Repertoires. https://doi.org/10.1101/2021.10.19.465057

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