Amperometric biosensor based on horseradish peroxidase immobilised magnetic microparticles
Amperometric biosensor based on horseradish peroxidase immobilised magnetic microparticles
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DOI：
10.1016/j.snb.2005.07.026
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发表时间：
2006-02
影响因子：
8.4
通讯作者：
Donghui Yu;B. Blankert;E. Bodoki;S. Bollo;J. Viré;R. Sandulescu;A. Nomura;J. Kauffmann
中科院分区：
文献类型：
--
作者：
Donghui Yu;B. Blankert;E. Bodoki;S. Bollo;J. Viré;R. Sandulescu;A. Nomura;J. Kauffmann
将具有高密度纳米孔的磁化硅基微粒(5μm)用于间歇条件下辣根过氧化物酶的固定化和电流型生物传感器的研制。该生物传感器被应用于研究氯氮平(CLZ)的过氧化作用，氯氮平是一种常用于治疗神经疾病的二苯并氮卓类药物。电流响应对应于CLZ氧化产物的电还原，即CLZ的氮离子和喹亚胺衍生物。尽管固定化辣根过氧化物酶的量相对较低(0.3molmolg)，但利用磁化的固体石蜡碳糊电极吸引微粒，实现了氯氮平在微摩尔浓度范围内的定量。底物的扩散和纳米孔内酶反应的产物被认为是生物传感器响应的限制因素。这种电流型生物传感器结构具有许多有趣的优点，如容易和可重复的微粒层更新，低酶消耗，可控的表面固定，保护酶微环境等。
Magnetised silica-based microparticles (MMPs) (5μm) with a high density of nanopores were used for horseradish peroxidase (HRP) immobilization and amperometric biosensor development in batch conditions. The resulting biosensor was applied to study the peroxidation of clozapine (CLZ) which is a dibenzoazepine drug often used in the treatment of neurological disorders. The amperometric response corresponded to the electroreduction of CLZ-oxidized products namely a nitrenium cation and quinoneimine derivative of CLZ. Despite a relatively low amount of immobilized HRP (0.3μmol/g), clozapine quantification in the micromolar concentration range was achieved by the use of a magnetized solid paraffin carbon paste electrode for microparticles attraction. Diffusion of substrate and products of the enzyme reaction within the nanopores were identified as limiting factors in the biosensor response. This amperometric biosensor configuration has number of interesting advantages such as ease and reproducible microparticle layer renewing, low enzyme consumption, controlled surface immobilization, protective enzyme microenvironment etc.