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KMID : 0385520050180060460
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Analytical Science & Technology
2005 Volume.18 No. 6 p.460 ~ p.468
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Characterization of biotin - avidin recognition system constructed on the solid substrate
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Lim JH
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Abstract
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The biotin-avidin complex, as a model recognition system, has been constructed through N-hydroxysuccinimide (NHS) reaction on a variety of substrates such as a smooth Au film, electrochemically roughened Au electrode and chemically modified mica. Stepwise self-assembled monolayers (SAMs) of biotin-avidin system were characterized by surface-enhanced resonance Raman scattering (SERRS) spectroscopy, atomic force microscopy (AFM) and surface plasmon resonance (SPR). A strong SERRS signal of rhodamine tags labeled in avidin from the SAMs on a roughened gold electrode indicated the successful complex formation of stepwise biotin-avidin recognition system. AFM images showed the circular shaped avidin aggregates (hexamer) with ca. 60 Å thick on the substrate, corresponding to one layer of avidin. The surface coverage and concentration of avidin molecules were estimated to be 90% and 7.5×10-¹² mol/㎠, respectively. SPR technique allowed one to monitor the surface reaction of the specific recognition with high sensitivity and precision.
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KEYWORD
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