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基于葡萄糖氧化酶催化金纳米粒子沉积的超灵敏电化学免疫传感器用于信号放大。

Ultrasensitive electrochemical immunosensor employing glucose oxidase catalyzed deposition of gold nanoparticles for signal amplification.

机构信息

Institute of Bioengineering and Nanotechnology, The Nanos, Singapore, Singapore.

出版信息

Biosens Bioelectron. 2011 Sep 15;27(1):53-7. doi: 10.1016/j.bios.2011.06.007. Epub 2011 Jun 17.

Abstract

This paper describes a novel enzymatic amplification strategy for ultrasensitive electrochemical immunosensing. This approach utilizes glucose oxidase for the enzymatic deposition of gold nanoparticles onto an indium tin oxide (ITO) electrode surface using a novel gold developer solution consisting of 20 mM of glucose, 20 mM of NaSCN, 0.5 M of p-benzoquinone (PBQ) and 1 mM of AuCl(4)(-) dissolved in 0.1 M of pH 7.5 phosphate buffer solution. The amount of gold deposited was quantified electrochemically by monitoring the reduction of gold oxide in an aqueous solution of 0.5 M of H(2)SO(4), which was correlated to the amount of antigens in the solution. The effectiveness of this strategy was demonstrated experimentally through the construction of an immunosensor for the detection of mouse IgG using a sandwich immunoassay in a linear dynamic range of 5 pg/ml to 50 ng/ml. A good mean apparent recovery in the range of 88-102% was obtained over the entire linear dynamic range of the sensor response in the serum samples. This suggested that the immunosensor would be useful for the testing of proteins in real clinical samples.

摘要

本文介绍了一种新颖的酶促扩增策略,用于超灵敏电化学免疫传感。该方法利用葡萄糖氧化酶,通过一种新型的金显影剂溶液,将金纳米粒子酶促沉积在氧化铟锡(ITO)电极表面上,该溶液由 20mM 的葡萄糖、20mM 的硫氰酸钠、0.5M 的对苯醌(PBQ)和 1mM 的 AuCl(4)(-)溶解在 0.1M pH7.5 的磷酸盐缓冲溶液中。通过监测 0.5M 的 H(2)SO(4)水溶液中氧化金的还原,电化学定量测定沉积的金量,这与溶液中抗原的量相关。通过使用夹心免疫分析法在血清样品的整个线性动态范围内,在 5pg/ml 至 50ng/ml 的线性动态范围内,对该策略的有效性进行了实验验证。在传感器响应的整个线性动态范围内,获得了 88-102%的良好平均表观回收率。这表明该免疫传感器将可用于实际临床样品中蛋白质的检测。

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