Electrochemical protein chip with arrayed immunosensors with antibodies immobilized in a plasma-polymerized film
Electrochemical protein chip with arrayed immunosensors with antibodies immobilized in a plasma-polymerized film
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DOI：
10.1021/ac0257391
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发表时间：
2003-03-01
影响因子：
7.4
通讯作者：
Karube, I
中科院分区：
文献类型：
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作者：
Kojima, K;Hiratsuka, A;Karube, I
制备了电化学蛋白质芯片。一个薄膜三电极系统，包括一个阵列的36个铂工作电极，一组薄膜Ag/AgCl电极，和铂辅助电极，集成在玻璃基板上。将捕获抗体固定在4.5 nm厚的双层六甲基二硅氧烷等离子体聚合膜中。由于其高度交联的网络结构，捕获抗体可以被牢固地固定。在一系列检测靶蛋白的过程中没有观察到非特异性吸附，相邻位点之间的电化学串扰可以忽略不计。使用模型蛋白、α-1-甲胎蛋白和β（2）-微球蛋白在单个芯片上进行夹心免疫测定。观察到葡萄糖氧化酶的酶促反应产生的过氧化氢氧化后的明显电流增加，这表明捕获蛋白实际上可以结合靶蛋白。用各自的捕获抗体在多个位点上独立地检测两种蛋白质。
An electrochemical protein chip was microfabricated. A thin-film three-electrode system, including an array of 36 platinum working electrodes, a set of thin-film Ag/AgCl electrodes, and platinum auxiliary electrodes, was integrated on a glass substrate. Capture antibodies were immobilized in a 4.5-nm-thick double layer of a hexamethyldisiloxane plasma-polymerized film. Because of their highly cross-linked network structure, the capture antibodies could be firmly immobilized. No nonspecific adsorption was observed during a series of procedures to detect target proteins, and electrochemical cross talk between neighboring sites was negligible. The sandwich immunoassay was conducted on a single chip using model proteins, alpha-1-fetoprotein and beta(2)-microglobulin. A distinct current increase following the oxidation of hydrogen peroxide produced by the enzymatic reaction of glucose oxidase was observed, which indicates that the capture proteins could actually bind the target proteins. Two kinds of protein were detected independently on multiple sites with respective capture antibodies.