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基于谷氨酸氧化酶固定在羧基化多壁碳纳米管/金纳米粒子/壳聚糖复合膜修饰金电极上的安培型谷氨酸生物传感器。

An amperometric glutamate biosensor based on immobilization of glutamate oxidase onto carboxylated multiwalled carbon nanotubes/gold nanoparticles/chitosan composite film modified Au electrode.

机构信息

Department of Biochemistry, M.D. University, Rohtak 124001, India.

出版信息

Biosens Bioelectron. 2013 Sep 15;47:496-501. doi: 10.1016/j.bios.2013.03.063. Epub 2013 Apr 2.

Abstract

A method is described for the construction of a novel amperometric glutamate biosensor based on covalent immobilization of glutamate oxidase (GluOx) onto, carboxylated multi walled carbon nanotubes (cMWCNT), gold nanoparticles (AuNPs) and chitosan (CHIT) composite film electrodeposited on the surface of a Au electrode. The GluOx/cMWCNT/AuNP/CHIT modified Au electrode was characterized by scanning electron microscopy (SEM), fourier transform infra-red (FTIR) spectroscopy, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The biosensor measured current due to electrons generated at 0.135V against Ag/AgCl from H2O2, which is produced from glutamate by immobilized GluOx. The biosensor showed optimum response within 2s at pH 7.5 and 35°C. A linear relationship was obtained between a wide glutamate concentration range (5-500μM) and current (μA) under optimum conditions. The biosensor showed high sensitivity (155nA/μM/cm(2)), low detection limit (1.6μM) and good storage stability. The biosensor was unaffected by a number of serum substances at their physiological concentrations. The biosensor was evaluated and employed for determination of glutamate in sera from apparently healthy subjects and persons suffering from epilepsy.

摘要

描述了一种基于共价固定化谷氨酸氧化酶(GluOx)到羧基化多壁碳纳米管(cMWCNT)、金纳米粒子(AuNPs)和壳聚糖(CHIT)复合膜的新型安培型谷氨酸生物传感器的构建方法,该复合膜沉积在 Au 电极表面。通过扫描电子显微镜(SEM)、傅里叶变换红外(FTIR)光谱、电化学阻抗谱(EIS)和循环伏安法(CV)对 GluOx/cMWCNT/AuNP/CHIT 修饰的 Au 电极进行了表征。该生物传感器测量了由于固定化 GluOx 产生的 H2O2 从 0.135V 对 Ag/AgCl 产生的电流。该生物传感器在 pH 7.5 和 35°C 下,在 2s 内达到最佳响应。在最佳条件下,在较宽的谷氨酸浓度范围内(5-500μM)与电流(μA)之间获得了线性关系。该生物传感器具有高灵敏度(155nA/μM/cm(2))、低检测限(1.6μM)和良好的储存稳定性。该生物传感器不受生理浓度下许多血清物质的影响。该生物传感器用于测定来自健康人和癫痫患者血清中的谷氨酸。

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