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

Fei Wen, Mason R. Smith, Huimin Zhao

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Antigenic peptides (termed T cell epitopes) are assembled with major histocompatibility complex (MHC) molecules and presented on the surface of antigen-presenting cells (APCs) for T cell recognition. T cells engage these peptide-MHCs using T cell receptors (TCRs). Because T cell epitopes determine the specificity of a T cell immune response, their prediction and identification are important steps in developing peptide-based vaccines and immunotherapies. In recent years, a number of computational methods have been developed to predict T cell epitopes by evaluating peptide-MHC binding; however, the success of these methods has been limited for MHC class II (MHCII) due to the structural complexity of MHCII antigen presentation. Moreover, while peptide-MHC binding is a prerequisite for a T cell epitope, it alone is not sufficient. Therefore, T cell epitope identification requires further functional verification of the MHC-binding peptide using 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 vector design, yeast surface display, peptide library generation in Escherichia coli, and functional screening in Saccharomyces cerevisiae are discussed.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages213-234
Number of pages22
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume2024
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CD4+ T cell epitope mapping
  • Flow cytometry
  • High-throughput screening
  • Human leukocyte antigen DR1 (HLA-DR1)
  • Influenza A virus
  • MHC-binding peptides
  • Major histocompatibility complex
  • Peptide library
  • Yeast display

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Construction and Screening of an Antigen-Derived Peptide Library Displayed on Yeast Cell Surface for CD4+ T Cell Epitope Identification'. Together they form a unique fingerprint.

Cite this