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Hypersensitive immunoassay by usingEscherichia coliouter membrane with autodisplayed Z-domains

Authors
Journal
Enzyme and Microbial Technology
0141-0229
Publisher
Elsevier
Publication Date
Volume
46
Identifiers
DOI: 10.1016/j.enzmictec.2009.12.004
Keywords
  • Immunoassay
  • Z-Domain
  • Orientation Control
  • C-Reactive Protein
  • Autodisplay
  • Outer Membrane
Disciplines
  • Biology
  • Medicine

Abstract

Abstract In this work, the Z-domain of protein A was autodisplayed on the surface of the Escherichia coli outer membrane as a fusion protein of AIDA-1, and then layered on a microplate in order to develop a hypersensitive immunoassay. By using microplates with controlled hydrophilicity the formation of outer membrane layer was carried out, and the driving force for the layer formation was proven to be a hydrophobic interactions. Through the orientation control of the immobilized antibodies by using autodisplayed Z-domains, the limit of detection (LOD) and the sensitivity of this new immunoassay were determined to have improved as much as 30-fold compared to the conventional ELISA. The applicability of the new immunoassay for medical diagnosis was demonstrated by the detection of C-reactive protein (CRP) which is known to be a biomarker protein for inflammatory diseases.

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