Garguilo M G, Huynh N, Proctor A, Michael A C
Department of Chemistry, University of Pittsburgh, Pennsylvania 15260.
Anal Chem. 1993 Mar 1;65(5):523-8. doi: 10.1021/ac00053a007.
Amperometric sensors have been developed for hydrogen peroxide, choline, and acetylcholine by immobilization of horseradish peroxidase, (HRP), choline oxidase, and acetylcholinesterase in a cross-linked redox polymer deposited on glassy carbon electrodes. Peroxide sensors, prepared by immobilization of HRP alone, gave detection limits of 10 nM and a linear response up to ca. 1 mM. Coimmobilization of HRP and glucose oxidase was used to establish the feasibility of highly efficient bienzyme sensors at low substrate levels. Replacing glucose oxidase with choline oxidase produced sensors with submicromolar detection limits and a linear response up to 0.8 mM. Addition of acetylcholinesterase to the sensors generated a relatively small response to acetylcholine that demonstrates the feasibility of trienzyme sensors. At low substrate concentrations, no loss in sensitivity during a 1-day experiment was observed. The response times of these sensors are all less than 30 s with 2-s response times achieved in some cases.
通过将辣根过氧化物酶（HRP）、胆碱氧化酶和乙酰胆碱酯酶固定在沉积于玻碳电极上的交联氧化还原聚合物中，已开发出用于过氧化氢、胆碱和乙酰胆碱的安培传感器。仅通过固定HRP制备的过氧化物传感器，检测限为10 nM，线性响应范围高达约1 mM。HRP和葡萄糖氧化酶的共固定用于确定高效双酶传感器在低底物水平下的可行性。用胆碱氧化酶替代葡萄糖氧化酶产生了检测限为亚微摩尔级且线性响应范围高达0.8 mM的传感器。向传感器中添加乙酰胆碱酯酶对乙酰胆碱产生了相对较小的响应，这证明了三酶传感器的可行性。在低底物浓度下，在为期1天的实验中未观察到灵敏度损失。这些传感器的响应时间均小于30秒，在某些情况下可达2秒。
