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Yeast Display for the Identification of Peptide-MHC Ligands of Immune Receptors

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Yeast Surface Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2491))

Abstract

T cells detect peptide antigens presented by major histocompatibility complex (MHC) proteins via their T cell receptor (TCR). The sequence diversity of possible antigens, with trillions of potential peptide-MHC targets, makes it challenging to study, characterize, and manipulate the peptide repertoire of a given TCR. Yeast display has been utilized to study the interactions between peptide-MHCs and T cell receptors to facilitate high-throughput screening of peptide-MHC libraries. Here we present insights on designing and validating a peptide-MHC yeast display construct, designing and constructing peptide libraries, conducting selections, and preparing, processing, and analyzing peptide library sequencing data. Applications for this approach are broad, including characterizing peptide-MHC recognition profiles for a TCR, screening for high-affinity mimotopes of known TCR-binding peptides, and identifying natural ligands of TCRs from expanded T cells.

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Acknowledgments

We would like to thank K. Christopher Garcia (Stanford University) for generous sharing of reagents and Christine Devlin for aiding in the creation of our yeast display protocols. This work was supported by National Science Foundation Graduate Research Fellowships to B.D.H, B.E.G., and P.V.H., and a Melanoma Research Alliance grant, the AACR-TESARO Career Development Award for Immuno-oncology Research (17-20-47-BIRN), Schmidt Futures, and the National Institutes of Health (P30CA14051 and 5U19AI110495) to M.E.B.

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Author notes

  1. Brooke D. Huisman and Beth E. Grace have contributed equally to this work.

Authors and Affiliations

  1. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA

    Brooke D. Huisman, Beth E. Grace, Patrick V. Holec & Michael E. Birnbaum

  2. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Brooke D. Huisman, Beth E. Grace, Patrick V. Holec & Michael E. Birnbaum

Authors
  1. Brooke D. Huisman
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  2. Beth E. Grace
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  3. Patrick V. Holec
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  4. Michael E. Birnbaum
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Corresponding author

Correspondence to Michael E. Birnbaum .

Editor information

Editors and Affiliations

  1. Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, Austria

    Michael W. Traxlmayr

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Huisman, B.D., Grace, B.E., Holec, P.V., Birnbaum, M.E. (2022). Yeast Display for the Identification of Peptide-MHC Ligands of Immune Receptors. In: Traxlmayr, M.W. (eds) Yeast Surface Display. Methods in Molecular Biology, vol 2491. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2285-8_15

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  • DOI: https://doi.org/10.1007/978-1-0716-2285-8_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2284-1

  • Online ISBN: 978-1-0716-2285-8

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