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Rapid Identification of MHCII-Binding Peptides Through Microsphere-Assisted Peptide Screening (MAPS)

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T-Cell Repertoire Characterization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2574))

Abstract

CD4+ T cells play a vital role in the immune response, and their function requires T cell receptor (TCR) recognition of peptide epitopes presented in complex with MHC class II (MHCII) molecules. Consequently, rapidly identifying peptides that bind MHCII is critical to understanding and treating infectious disease, cancer, autoimmunity, allergy, and transplant rejection. Computational methods provide a fast, ultrahigh-throughput approach to predict MHCII-binding peptides but lack the accuracy of experimental methods. In contrast, experimental methods offer accurate, quantitative results at the expense of speed. To address the gap between these two approaches, we developed a high-throughput, semiquantitative experimental screening strategy termed microsphere-assisted peptide screening (MAPS). Here, we use the Zika virus envelope protein as an example to demonstrate the rapid identification of MHCII-binding peptides from a single pathogenic protein using MAPS. This process involves several key steps including peptide library design, peptide exchange into MHCII, peptide-MHCII loading onto microspheres, flow cytometry screening, and data analysis to identify peptides that bind to one or more MHCII alleles.

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Author information

Author notes

  1. Luke F. Bugada and Mason R. Smith contributed equally.

Authors and Affiliations

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

    Luke F. Bugada, Mason R. Smith & Fei Wen

Authors
  1. Luke F. Bugada
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  2. Mason R. Smith
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  3. Fei Wen
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Corresponding author

Correspondence to Fei Wen .

Editor information

Editors and Affiliations

  1. Institute of Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA, USA

    Huang Huang

  2. Institute of Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA, USA

    Mark M. Davis

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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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Bugada, L.F., Smith, M.R., Wen, F. (2022). Rapid Identification of MHCII-Binding Peptides Through Microsphere-Assisted Peptide Screening (MAPS). In: Huang, H., Davis, M.M. (eds) T-Cell Repertoire Characterization. Methods in Molecular Biology, vol 2574. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2712-9_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2712-9_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2711-2

  • Online ISBN: 978-1-0716-2712-9

  • eBook Packages: Springer Protocols

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