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Australian Journal of Chemistry Australian Journal of Chemistry Society
An international journal for chemical science
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Avidin–Biotin Assembling of Horseradish Peroxidase Multi-Monomolecular Layers on Electrodes

Benoît Limoges A B , Jean-Michel Savéant A B and Dounia Yazidi A
+ Author Affiliations
- Author Affiliations

A Laboratoire d’Electrochimie Moléculaire, UMR CNRS 7591, Université de Paris 7 ‘Denis Diderot’, 75251 Paris Cedex 05, France.

B Corresponding authors. Email: limoges@paris7.jussieu.fr; saveant@paris7.jussieu.fr

Australian Journal of Chemistry 59(4) 257-259 https://doi.org/10.1071/CH06039
Submitted: 6 February 2006  Accepted: 23 March 2006   Published: 1 May 2006

Abstract

The strong avidin–biotin affinity is used to stack up successive monomolecular layers of horseradish peroxidase on carbon electrodes. After a biotinylated immunoglobulin is adsorbed on the electrode surface, alternate deposition of neutravidin and biotinylated HRP allows the assemblage of up to 16 successive active HRP layers. The film build-up is followed by cyclic voltammetry using an osmium complex as soluble mediator and H2O2 as substrate. The variation of the resulting catalytic responses with H2O2 concentration exhibit characteristics qualitatively consistent with the catalysis-inhibition reaction scheme previously established for monomolecular layers. In most cases the catalytic activity increases steadily with the number of monomolecular layers, leading to a significant increase of the analytical sensitivity of the derivatized electrode.


References


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        | Crossref |  GoogleScholarGoogle Scholar |  that a cosubstrate concentration as low as 0.2 µM can be easily determined. With a background current of 1.5 nA at 10 mV s−1 and a signal-to-noise ratio of 3, this means that the detection limit is as low as 1 nM.