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Construction and Screening of an Antigen-Derived Peptide Library Displayed on Yeast Cell Surface for CD4+ T Cell Epitope Identification

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Immunoproteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1061))

Abstract

Identification of T cell epitopes is a critical, but often difficult step in studying T cell function and developing peptide-based vaccines and immunotherapies. Unlike antibodies that recognize free soluble antigens, T cell receptor (TCR) recognizes its epitope bound to major histocompatibility complex (MHC) expressed on antigen presenting cells (APCs). In addition, the examination of T cell epitope activity requires the use of professional APCs, which are difficult to isolate, expand, and maintain. To address these issues, we have developed a facile, accurate, and high-throughput method for T cell epitope mapping by screening antigen-derived peptide libraries in complex with MHC protein displayed on yeast cell surface. Here, we use hemagglutinin and influenza A virus X31/A/Aichi/68 as examples to describe the key steps in identification of CD4+ T cell epitopes from a single antigenic protein and the entire genome of a pathogen, respectively. Methods for single-chain peptide-MHC complex vector design, yeast surface display, peptide library generation in Escherichia coli, and functional screening in Saccharomyces cerevisiae are discussed.

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Author information

Authors and Affiliations

  1. Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA

    Fei Wen

  2. Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Huimin Zhao

  3. Departments of Chemistry, Biochemistry, and Bioengineering, Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Huimin Zhao

Authors
  1. Fei Wen
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  2. Huimin Zhao
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Editor information

Editors and Affiliations

  1. National Research Council Canada Human Health Therapeutics, Ottawa, Ontario, Canada

    Kelly M. Fulton

  2. National Research Council Canada Human Health Therapeutics, Ottawa, Ontario, Canada

    Susan M. Twine

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Wen, F., Zhao, H. (2013). Construction and Screening of an Antigen-Derived Peptide Library Displayed on Yeast Cell Surface for CD4+ T Cell Epitope Identification. In: Fulton, K., Twine, S. (eds) Immunoproteomics. Methods in Molecular Biology, vol 1061. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-589-7_15

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  • DOI: https://doi.org/10.1007/978-1-62703-589-7_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-588-0

  • Online ISBN: 978-1-62703-589-7

  • eBook Packages: Springer Protocols

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