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KMID : 0545120020120050780
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Journal of Microbiology and Biotechnology
2002 Volume.12 No. 5 p.780 ~ p.786
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Detection of Escherichia coli 0157:H7 Using Immunosensor Based on Surface Plasmon Resonance
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OH, BYUNG-KEUN
KIM, YOUNG-KEE/BAE, YOUNG MIN/LEE, WON HONG/CHOI, JEONG-WOO
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Abstract
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An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Esherichia coli O157:H7. A self-assembled protein G layer in 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 (EDZC) 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. coli 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.
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