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- E-ISSN 2288-8985
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Characterization of biotin-avidin recognition system constructed on the solid substrate
Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2005, v.18 no.6, pp.460-468
(2005). Characterization of biotin-avidin recognition system constructed on the solid substrate. Analytical Science and Technology, 18(6), 460-468.
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Abstract
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. <TEX>$60{\AA}$</TEX> thick on the substrate, corresponding to one layer of avidin. The surface coverage and concentration of avidin molecules were estimated to be 90% and <TEX>$7.5{\times}10^{-12}mol/cm^2$</TEX>, respectively. SPR technique allowed one to monitor the surface reaction of the specific recognition with high sensitivity and precision.
- keywords
- Biotin, avidin, stepwise self-assembly, SERRS, AFM, SPR
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