Publication
602
J. Am. Chem. Soc. , 128, 6014-6015, 2006.
DOI: 10.1021/ja060801n
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High Amplification Rates from the Association of Two Enzymes
Confined within a Nanometric Layer Immobilized on an Electrode: Modeling and
Illustrating Example
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Benoît Limoges*, François Mavré, Damien Marchal, and Jean-Michel Savéant*
Contribution from the Laboratoire d'Electrochimie Moléculaire, Université de Paris 7-Denis Diderot, 2 place Jussieu,75251 Paris Cedex 05, France
Electrochemical responses (e.g., chronoamperometric) obtained with an
immobilized enzyme that produces an electroactive species may be used to
quantitate the amount of enzyme or the concentration of its substrate. It is
shown, on theoretical and experimental bases, that product-to-substrate coupling
with a second enzyme co-immobilized with the first within one or within a small
number of monolayers, allows high amplification rates (higher than 1000), avoids
membrane transport limitations, and lends itself to precise kinetic analyses
that provide guidelines for optimization of the analytical sensitivity. Very
large amplification factors, as large as several thousands, can be reached
experimentally, in agreement with appropriately derived theoretical predictions,
thus opening the route to the rational design of high-performance substrate
sensing or affinity assays applications. |