电化学生物传感器 2012

An aqueous media based approach for the preparation of a biosensor platform composed of graphene oxide and Pt-black.

Biosensors & bioelectronics Shi J, Zhang H, Snyder A, Wang MX, Xie J, Marshall Porterfield D, Stanciu LA
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组成图示

An aqueous media based approach for t... 传感器构成示意图

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传感器类型

电化学生物传感器

检测对象

葡萄糖(Glucose/D-glucose);样品基质:pH 7.4 PBS 混合溶液(含人血常见干扰物测试)

检测原理

该传感器为酶促电化学检测。固定化葡萄糖氧化酶(GOx)特异性催化葡萄糖与氧气反应生成葡萄糖酸和过氧化氢(H2O2)。在 +500 mV 工作电位下,H2O2 在 Pt-black/GrOx 复合微电极表面发生电催化氧化生成氧气,释放电子并经 Pt/Ir 微电极形成与葡萄糖浓度成正比的安培电流。GrOx 作为水溶性纳米模板引导 Pt-black 沿其生长,形成高比表面积纳米图案,提高有效表面积和 H2O2 氧化电催化活性,同时促进电子转移并降低传质限制,从而实现信号放大。

检测灵敏度

LOD: 1 mM;线性范围: 1 mM–2 mM;R = 0.999;灵敏度: 465.9±48.0 nA/mM;响应时间: ~4 s

效应效果

GrOx/Pt-black 葡萄糖微传感器对常见人血干扰物选择性良好:2 mM 葡萄糖下,125 μM 抗坏血酸、330 μM 尿酸、130 μM 对乙酰氨基酚的电流响应分别仅为葡萄糖的 0.2%、4.8%、0.8%。重复测试灵敏度变化 4.8±0.2%;-20 °C 空气保存 7 天灵敏度下降 9%。与 Pt-black 相比,葡萄糖灵敏度提高 25 倍以上,H2O2 电催化灵敏度提高 12 倍以上,有效表面积约为其 12 倍。表 1 显示其灵敏度密度在所列工作中最高,但线性范围不及部分报道。作者认为可用于癌症细胞葡萄糖代谢研究。

传感器的构成

  • 基底/换能器电极:Pt/Ir 微电极(Pt/Ir wire microelectrode,PI20033.0A10,线径 0.256 mm,尖端 1–2 mm),作为工作微电极和电子传导基底。
  • 纳米模板修饰层:水溶性氧化石墨烯(GrOx,graphene oxide)滴涂风干,作为 Pt-black 电沉积的纳米分子模板并增加有效表面积。
  • 电催化修饰层:铂黑(Pt-black,无定形铂纳米团簇)在 GrOx 上电沉积,提供 H2O2 氧化电催化活性并包裹 GrOx 防止溶出。
  • 识别元件:葡萄糖氧化酶(GOx,glucose oxidase,E.C.1.1.3.4)催化葡萄糖氧化生成 H2O2。
  • 交联固定剂:戊二醛(glutaraldehyde,2.5% 水溶液)通过席夫碱共价交联 GOx 至电极表面。
  • 封闭/保护剂:牛血清白蛋白(BSA,bovine serum albumin)保护酶并辅助固定。

中文摘要

本文报道了一种基于水相制备氧化石墨烯(GrOx)/铂黑(Pt-black)纳米复合生物传感器平台的方法。利用水溶性 GrOx 与石墨烯结构相似的特点,将 GrOx 滴涂于微电极表面作为纳米分子模板,再电沉积无定形铂纳米团簇 Pt-black,使 Pt-black 沿 GrOx 生长。扫描电镜和能谱显示 Pt-black 沿 GrOx 分布,复合微电极的有效表面积和对 H2O2 氧化的电催化活性显著高于单独 Pt-black 微电极。通过戊二醛交联固定葡萄糖氧化酶(GOx)制备葡萄糖生物纳米复合微传感器后,GrOx/Pt-black 微传感器的灵敏度高于 Pt-black 微传感器,表明该纳米复合结构促进了电子转移和/或降低了传质限制。所得葡萄糖微传感器灵敏度为 465.9±48.0 nA/mM,检出限为 1 mM,线性范围为 1 mM–2 mM,响应时间约 4 s,并具有良好的稳定性和对常见干扰物的选择性。

英文摘要

The combination of Pt nanoparticles and graphene was more effective in enhancing biosensing than either nanomaterial alone according to previous reports. Based on the structural similarities between water soluble graphene oxide (GrO(x)) and graphene, we report the fabrication of an aqueous media based GrO(x)/Pt-black nanocomposite for biosensing enhancement. In this approach GrO(x) acted as a nanoscale molecular template for the electrodeposition of Pt-black, an amorphously nanopatterned isoform of platinum metal. Scanning electron microscopy (SEM) images and energy-dispersive X-ray spectroscopy (EDS) showed that Pt-black was growing along GrO(x). The effective surface area and electrocatalytic activity towards H(2)O(2) oxidation of GrO(x)/Pt-black microelectrodes were significantly higher than for Pt-black microelectrodes. When used to prepare a bio-nanocomposite based on protein functionalization with the enzyme glucose oxidase (GOx), the GrO(x)/Pt-black microbiosensors exhibited improved sensitivity over the Pt-black microbiosensors. This suggested that the GrO(x)/Pt-black nanocomposite facilitated an increase in electron transfer, and/or minimized mass transport limitations as compared to Pt-black used alone. Glucose microbiosensors based on GrO(x)/Pt-black exhibited high sensitivity (465.9 ± 48.0 nA/mM), a low detection limit of 1 μM, a linear response range of 1 μM-2mM, and response time of ≈ 4s. Additionally the sensor was stable and highly selective over potential interferents.