Limoges Benoît, Marchal Damien, Mavré Francois, Savéant Jean-Michel
Laboratoire d'Electrochimie Moléculaire, UMR CNRS 7591, Université de Paris 7 - Denis Diderot, 2 place Jussieu, 75251 Paris Cedex 05, France.
J Am Chem Soc. 2006 May 10;128(18):6014-5. doi: 10.1021/ja060801n.
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.
用产生电活性物质的固定化酶获得的电化学响应（如计时电流法）可用于定量酶的量或其底物的浓度。理论和实验表明，与第一种酶共固定在一个或少数几个单层内的第二种酶实现产物与底物的偶联，可实现高放大率（高于1000），避免膜传输限制，并适用于精确的动力学分析，为优化分析灵敏度提供指导。通过实验可以达到高达数千的非常大的放大因子，这与适当推导的理论预测一致，从而为高性能底物传感或亲和分析应用的合理设计开辟了道路。
