Identifying Bacterial Immune Evasion Proteins Using Phage Display

Methods Mol Biol. 2017;1535:43-61. doi: 10.1007/978-1-4939-6673-8_4.

Abstract

Methods aimed at identification of immune evasion proteins are mainly rely on in silico prediction of sequence, structural homology to known evasion proteins or use a proteomics driven approach. Although proven successful these methods are limited by a low efficiency and or lack of functional identification. Here we describe a high-throughput genomic strategy to functionally identify bacterial immune evasion proteins using phage display technology. Genomic bacterial DNA is randomly fragmented and ligated into a phage display vector that is used to create a phage display library expressing bacterial secreted and membrane bound proteins. This library is used to select displayed bacterial secretome proteins that interact with host immune components.

Keywords: Functional identification; High-throughput; Immune evasion; Phage display; Secretome.

MeSH terms

  • Bacteria / immunology*
  • Bacteria / metabolism*
  • Bacteria / pathogenicity
  • Bacterial Proteins / immunology*
  • Bacterial Proteins / metabolism*
  • Cell Surface Display Techniques*
  • Genomic Library
  • High-Throughput Screening Assays
  • Immune Evasion*
  • Peptide Library
  • Proteome
  • Proteomics / methods

Substances

  • Bacterial Proteins
  • Peptide Library
  • Proteome