Construction and screening of an antigen-derived peptide library displayed on yeast cell surface for CD4+ T cell epitope identification

Fei Wen, Huimin Zhao

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationImmunoproteomics
Subtitle of host publicationMethods and Protocols
EditorsKelly Fulton, Susan Twine
Pages245-264
Number of pages20
DOIs
StatePublished - Sep 20 2013

Publication series

NameMethods in Molecular Biology
Volume1061
ISSN (Print)1064-3745

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Keywords

  • CD4 T cell epitope mapping
  • Flow cytometry
  • High throughput screening
  • HLA-DR1
  • Influenza A virus
  • Major histocompatibility complex
  • Peptide library
  • Single-chain peptide-MHC complex
  • Yeast display

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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 K. Fulton, & S. Twine (Eds.), Immunoproteomics: Methods and Protocols (pp. 245-264). (Methods in Molecular Biology; Vol. 1061). https://doi.org/10.1007/978-1-62703-589-7_15