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Pre-absorbed Immunoproteomics: A Novel Method for the Detection of Bacterial Surface Proteins

  • Guangjin Liu
  • Wei Zhang
  • Chengping Lu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1061)

Abstract

Bacterial surface proteins are often investigated as potential vaccine candidates and biomarkers of virulence. In this chapter, a novel method for identifying bacterial surface proteins is presented, which combines immunoproteomic with immunoserologic techniques. Immunoproteomics, involving the separation of proteins by two-dimensional electrophoresis (2-DE) and Western blotting, has become an increasingly popular method for identifying immunoreactive proteins. In conventional serological technique, cross-absorption is a powerful method used to minimize cross reaction during agglutination assays. The serum pre-absorption process in our method was developed from cross-absorption but modified to remove antibodies that recognized bacterial surface antigens, thereby generating pre-absorbed sera. These pre-absorbed sera were used in Western blotting after 2-DE to find bacterial surface protein antigens. This new method has been proven to be a useful tool for identifying surface proteins, and aid in the development of new vaccine subunits and disease diagnostics.

Key words

Bacteria Immunoproteomic Immunoserology Pre-absorbed 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Guangjin Liu
    • 1
  • Wei Zhang
    • 1
  • Chengping Lu
    • 1
  1. 1.Key Laboratory of Animal Bacteriology, Ministry of AgricultureNanjing Agricultural UniversityNanjingChina

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