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Preparation of porous titania sol-gel matrix for immobilization of horseradish peroxidase by a vapor deposition method

机译:气相沉积法制备用于固定辣根过氧化物酶的多孔二氧化钛溶胶-凝胶基质

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摘要

A new and facile vapor deposition method has been developed for the preparation of sol-gel matrix. This method was used to form a titania sol-gel thin film and to immobilize horseradish peroxidase (HRP) on a glassy carbon electrode surface for the production of an ampero-metric hydrogen peroxide biosensor. This process prevented the cracking of conventional sol-gel-detived glasses. The morphologies of both titania sol-gel and the enzyme membranes were characterized using scanning electron microscopy and proved to be chemically clean, porous, and homogeneous and to have a very narrow particle size distribution. The sol-gel-derived titania-modified electrode retained the enzyme bioactivity and provided for long-term stability of the enzyme in storage. In the presence of catechol as a mediator, the sensor exhibited a rapid electrocatalytic response (less than 5 s), a linear calibration range from 0.08 to 0.56 mM with a detection limit of 1.5 muM and a high sensitivity (61.5 muA mM(-1)) for monitoring of H2O2. Effects of pH and operating potential were also explored for optimum analytical performance by using the amperometric method. The apparent Michaelis-Menten constant of the encapsulated HRP was 1.89 +/- 0.21 mM. [References: 39]
机译:已经开发了一种新的且容易的气相沉积方法来制备溶胶-凝胶基质。该方法用于形成二氧化钛溶胶-凝胶薄膜,并将辣根过氧化物酶(HRP)固定在玻璃碳电极表面上,用于生产安培型过氧化氢生物传感器。这个过程防止了传统的溶胶凝胶玻璃破裂。使用扫描电子显微镜对二氧化钛溶胶-凝胶和酶膜的形态进行了表征,并证明其化学清洁,多孔和均质,并且具有非常窄的粒度分布。溶胶-凝胶衍生的二氧化钛修饰的电极保留了酶的生物活性,并提供了酶在储存中的长期稳定性。在存在儿茶酚作为介质的情况下,传感器表现出快速的电催化响应(小于5 s),线性校准范围为0.08至0.56 mM,检测限为1.5μM和高灵敏度(61.5μAmM(-1) ))用于监测H2O2。还通过使用安培法探索了pH和操作电位的影响,以获得最佳分析性能。包封的HRP的表观Michaelis-Menten常数为1.89 +/- 0.21mM。 [参考:39]

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