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Related Experiment Video

Updated: Jun 13, 2025

Protein Engineering by Yeast Surface Display
05:49

Protein Engineering by Yeast Surface Display

Published on: November 29, 2024

1.1K

High-throughput screening for class I peptide MHC binding via yeast surface display.

Patrick V Holec, Kathryn C Breuckman, Owen Leddy

    Biorxiv : the Preprint Server for Biology
    |June 12, 2025
    PubMed
    Summary

    This study introduces a novel yeast display pipeline for rapidly identifying peptide-MHC class I binders across diverse alleles, aiding in understanding adaptive immunity and drug hypersensitivity.

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    Area of Science:

    • Immunology
    • Molecular Biology
    • Proteomics

    Background:

    • T cells are crucial for adaptive immunity, recognizing short peptides presented by Major Histocompatibility Complex (MHC) molecules.
    • Evaluating stable peptide presentation across diverse MHC alleles is challenging due to the high polymorphism of MHCs.
    • Existing methods lack the systemic evaluation capability for peptide-MHC (pMHC) interactions across a broad range of MHC alleles.

    Purpose of the Study:

    • To develop and present a yeast display pipeline for high-throughput screening of peptide-MHC class I (pMHC-I) binders.
    • To enable rapid identification of potential T cell antigens from large proteomic datasets.
    • To investigate biological phenomena like drug-induced hypersensitivity impacting peptide presentation.

    Main Methods:

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    Last Updated: Jun 13, 2025

    Protein Engineering by Yeast Surface Display
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    • A yeast display pipeline was engineered for large-scale screening of peptide-MHC class I binders.
    • The pipeline was applied to screen proteomic data from various pathogens, including Mtb, SARS-CoV-2, Dengue, and Zika viruses.
    • The system allows for the evaluation of peptide binding across numerous MHC alleles simultaneously.

    Main Results:

    • The yeast display pipeline successfully identified pMHC-I binders across multiple MHC alleles.
    • The study captured novel biological insights, such as interference with peptide presentation in type IV drug-induced hypersensitivity.
    • A high-resolution catalog of potential T cell antigens from diverse pathogen proteomes was generated.

    Conclusions:

    • The developed yeast display platform offers a rapid and scalable method for generating unbiased datasets of pMHC-I interactions.
    • This tool facilitates the discovery of T cell antigens and enhances the understanding of immune responses.
    • The platform provides a flexible approach to study peptide presentation relevant to infectious diseases and drug hypersensitivity.