辣根过氧化物酶在巯基丁二酸铜(II)自组装单分子层修饰电极表面的固定及对H_2O_2传感研究  

Incorporation of Horseradish Peroxidase on Self-Assembled Monolayer Modified Electrode by Mercapto-Butanedioic Acid Combined Copper(II) and Study of Sensitivity to H_2O_2

作　　者：干宁[1] 余杨锋[1] 王志颖[1] 葛从辛[1] 徐伟民[1] 

机构地区：[1]宁波市新型功能材料及其制备科学国家重点实验室宁波大学材料与化工学院

出　　处：《传感技术学报》2007年第2期262-266,共5页Chinese Journal of Sensors and Actuators

基　　金：国家自然科学基金资助(60508012);浙江省科技攻关项目资助(2006C31040);浙江省自然科学基金资助(Y106725)

摘　　要：利用气相沉积技术制备TiO2凝胶膜,将辣根过氧化酶固定在巯基丁二酸铜(II)(CuL-SH)自组装单分子层修饰金电极表面,制得测定H2O2的电流型生物传感器.金电极表面的CuL-SH同时作为电子媒介体.研究了各种因素如pH、工作电位等对传感器响应电流的影响.传感器对H2O2的还原显示出快速电催化响应(<10s),计时电流法测定H2O2的线性范围为2.2μmol/L-0.6mmol/L(R=0.999),检出限为1.0×10-6mol/L.测得酶催化动力学参数米氏常数KMapp=0.56mmol/L.传感器的稳定性好,60d其响应值仍保持90%.对其灵敏度和选择性进行了研究,并应用于实际样品测定.A vapor depositive titania （TiO2） sol-gel film was developed for the fabrication of an amperometric hydrogen peroxide （H2O2） biosensor. The film was used to immobilize horseradish peroxidase （HRP） on a modified Au electrode self-assembled by Mercapto-butanedioic acid combined copper （Ⅱ） （CuL-SH） monolayer which was also employed as an electron mediator. Effects of some experimental conditions such as pH , applied potential on the response current of the biosensor were investigated. The biosensor had a fast response of less than 10 s and linear calibration range of 2.2×10^-6-6. 0 × 10^-4 mol/L with a correlation coefficient of 0. 998 9 and detection limit of 1. 0× 10^-6 mol/L for H2O2. The Michaelis-Menten constant KMapp was found to be 0. 56 mmol/ L. The sensor remains 90 % of its response after using 60 days. Both sensitivity and selectivity of the biosensor were investigated. This method was used to determine the content of H2O2 in the real sample.

关 键 词：过氧化氢生物传感器 气相沉积 二氧化钛溶胶-凝胶 巯基丁二酸铜(Ⅱ) 辣根过氧化物酶 自组装单分子层 电催化 

分 类 号：O657.1[理学—分析化学]