High-throughput t-cell epitope discovery through mhc peptide exchange

Sine Reker Hadrup, Mireille Toebes, Boris Rodenko, Arnold H. Bakker, David A. Egan, Huib Ovaa, Ton N. M. Schumacher

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

Recombinant major histocompatibility complex (MHC) class I molecules complexed with pathogen-specific or other disease-associated antigens have become essential reagents for the analysis of adaptive T-cell responses. However, conventional techniques for the production of recombinant peptide-MHC (pMHC) complexes are highly involved and thereby limit the use of pMHC complexes in terms of antigen diversity. To make pMHC-based techniques suitable for high-throughput analyses we developed an MHC peptide exchange technology based on the use of conditional MHC ligands. This technology enables the parallel production of thousands of different pMHC complexes within hours, allowing the development of high-throughput MHC-based assay systems to identify MHC ligands and cytotoxic T-cell responses. These high-throughput assays should prove valuable for the screening of entire disease-associated proteomes, including pathogen-encoded proteomes, tumor-associated antigens, and autoimmune antigens. �� 2009 Humana Press, a part of Springer Science+Business Media, LLC.
Original languageEnglish
Title of host publicationEpitope Mapping Protocols
Number of pages23
Volume524
PublisherHumana Press
Publication date2009
Edition2
Pages383-405
DOIs
Publication statusPublished - 2009
Externally publishedYes
SeriesMethods in Molecular Biology
ISSN1064-3745

Keywords

  • Genetics
  • Molecular Biology
  • CD8+ T cells
  • Conditional ligands
  • High-throughput screening
  • MHC peptide exchange
  • T-cell epitopes
  • epitope
  • ligand
  • peptide
  • peptide library
  • article
  • biotinylation
  • CD8+ T lymphocyte
  • economics
  • epitope mapping
  • gene
  • genetics
  • human
  • immunology
  • methodology
  • protein binding
  • protein folding
  • synthesis
  • ultraviolet radiation
  • Biotinylation
  • CD8-Positive T-Lymphocytes
  • Epitope Mapping
  • Epitopes, T-Lymphocyte
  • Genes, MHC Class I
  • Humans
  • Ligands
  • Peptide Library
  • Peptides
  • Protein Binding
  • Protein Folding
  • Ultraviolet Rays
  • major histocompatibility complex peptide MHC peptide
  • T-cell epitope
  • 02506, Cytology - Animal
  • 10060, Biochemistry studies - General
  • 15002, Blood - Blood and lymph studies
  • 15004, Blood - Blood cell studies
  • 34502, Immunology - General and methods
  • Chemical Coordination and Homeostasis
  • cytotoxic T-cell immune system, blood and lymphatics
  • major histocompatibility complex peptide exchange technique laboratory techniques, immunologic techniques
  • Biochemistry and Molecular Biophysics
  • Immune System
  • Methods and Techniques

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