传感器类型
电化学生物传感器
检测对象
儿茶酚(catechol);样品基质为磷酸盐缓冲液(PBS,含0.1 mM H2O2),面向环境水样应用。
检测原理
该传感器以HRP为生物识别/催化元件。在固定电位−0.1 V(vs Ag/AgCl)下,HRP催化H2O2还原为水,同时将儿茶酚氧化为邻苯醌。邻苯醌在电极表面被还原回儿茶酚,产生阴极安培电流。儿茶酚浓度升高时,酶促氧化通量和界面还原电流随之增大,形成线性响应。GNP修饰显著增大电极微观有效面积并改善电子传递,UMEA提供高电流密度与快速传质,二者共同放大信号。
检测灵敏度
LOD: 0.05 mM;线性范围: 0.1 mM–0.4 mM;灵敏度: 228.6 μA cm−2 mM−1;R^2 = 0.9912
效应效果
与裸UMEA和微电极生物传感器相比,GNP修饰UMEA的灵敏度分别提高约3倍和80倍。三种构型的重现性RSD均低于10%,单器件五次校准重复性RSD低于6%。GNP/UMEA在0.1–0.4 mM范围内线性,LOD为0.05 mM;微电极LOD为0.015 mM。论文未报告实际样品加标回收率、选择性或抗干扰实验。作者认为该平台可缩小至平面超微电极阵列,适用于环境酚类检测,并有望用于自动化和多路检测。
传感器的构成
- 基底/换能器电极:金超微电极阵列(Au UMEA),光刻制备,作为安培检测工作电极;片上金对电极(Au CE)用于三电极体系
- 纳米材料修饰层:金纳米粒子(GNP),电沉积于 UMEA 表面,增大微观有效面积并保持电极特性
- 自组装单分子层:双硫代双-N-琥珀酰亚胺基丙酸酯(DTSP)硫醇 SAM,锚定金/GNP 表面并提供琥珀酰亚胺反应位点
- 识别元件:辣根过氧化物酶(HRP),通过肽键共价固定于 DTSP,催化 H2O2 还原和儿茶酚氧化
- 信号底物/电子供体:过氧化氢(H2O2)与儿茶酚(catechol),H2O2 为酶促电子受体,儿茶酚为目标物兼电子供体
- 清洗/封闭:Tween 20 清洗,去除非特异性吸附的 HRP
- 缓冲介质:磷酸盐缓冲液(PBS,pH 7.4),维持酶活性并提供检测介质
中文摘要
报道了金纳米粒子修饰的超微电极阵列(GNP/UMEA)作为生物传感器换能平台的比较评估。将金纳米粒子电沉积于金超微电极阵列表面,可在不改变其固有电极特性的前提下将有效面积提高约百倍。以辣根过氧化物酶(HRP)为识别元件,通过硫醇自组装单分子层(SAM)共价固定于裸金微电极、裸金超微电极阵列及金纳米粒子修饰超微电极阵列三种换能平台上。所得生物传感器在固定电位−0.1 V(vs Ag/AgCl)下用于儿茶酚(catechol)的安培检测。与裸超微电极阵列和微电极生物传感器相比,金纳米粒子修饰超微电极阵列使灵敏度分别提高约3倍和80倍。该传感器对儿茶酚在0.1–0.4 mM范围内呈线性响应,检出限为0.05 mM。结果证明金纳米粒子修饰超微电极阵列是构建高灵敏度生物传感器的优良平台。
英文摘要
Gold ultramicroelectrode arrays (UMEAs) modified with gold nanoparticles (GNP) are shown to be a highly suitable transducer platform for the fabrication of biosensors. Comparative studies were carried out with microelectrodes and UMEAs, the latter being either bare or modified with GNPs. GNPs could be electrodeposited on to the UMEA surface, thereby increasing its active area up to one hundred times but without affecting its inherent electrodic properties. Horseradish peroxidase enzyme (HRP) was covalently immobilized over the three different transducer platforms by means of a thiol self-assembled monolayer (SAM). The resulting biosensors were applied to the amperometric detection of catechol, selected as a target analyte, at a set potential of -0.1 V vs. Ag/AgCl. The use of GNP-modified UMEAs increased the sensitivity of the developed biosensor 3-fold and 80-fold compared with the values achieved with bare UMEA and microelectrode based biosensors, respectively. The GNP-modified UMEA based biosensor showed a linear response to catechol in the concentration range from 0.1 mM to 0.4 mM, with a limit of detection of 0.05 mM.