Detection of Escherichia coli O157:H7 Using Immunosensor Based on Surface Plasmon Resonance

  • Oh, Byung-Keun (Department of Chemical Engineering, Sogang University) ;
  • Kim, Young-Kee (Department of Chemical Engineering, Hankyong National University) ;
  • Bae, Young-Min (Department of Chemical Engineering, Sogang University) ;
  • Lee, Won-Hong (Department of Chemical Engineering, Sogang University) ;
  • Choi, Jeong-Woo (Department of Chemical Engineering, Sogang University)
  • Published : 2002.10.01

Abstract

An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

Keywords

References

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