Development of highly integrated enzyme switches as a multi-parameter sensing device

开发高度集成的酶开关作为多参数传感装置

基本信息

批准号：
14550779
负责人：
SUZUKI Masayasu
金额：
$ 2.24万
依托单位：
University of Toyama
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14550779/
关键词：
Biosensor Enzyme sensor Enzyme switch Poly(aniline)Integration Glucose sensor Lactate sensor

项目摘要

Enzyme switch is a novel biosensing device based on the conductometric measurements of polyaniline(PAn) film covered with enzymes. The enzyme switch might have following advantages over conventional enzyme sensors : 1)Sensing chip itself has a memory function, 2)response might be independent from the electrode size, 3)Miniaturization and integration are easy because of its simple structure. 4)Easy connection with actuators, since the resistance change is index. In this study, these advantages were verified by using prepared enzyme switches.PAn loses its electroconductivity by oxidation and recovers it by reduction. If the oxidase-peroxidase(HRP) co-immobilized electrodes are prepared with PAn, when the produced compounds I and II are reduced at the surface of PAn,PAn might be oxidized and may lose its conductivity. By monitoring conductivity of PAn, "enzyme-switch", which gives "off" signal by the existence of specific substrate, might be constructed.PAn film was prepared electrochemically over the two micro-patterned thin film electrodes under the acidic conditions. Then glucose oxidase(or lactate oxidase) and HRP were immobilized onto the PAn film by glutaraldehyde crosslinking method. By the addition of substrates, the decrease of PAn conductivity due to the enzyme reaction could be observed. The oxidized PAn recovered its conductivity by applying 0.1 V vs.Ag/AgCl. In flow system, it took approximately 3 min for switching from "on" to "off", or "off" to "on", and peak heights were correspond to substrate concentration. The prepared enzyme switch had advantages both as enzyme test strips and enzyme sensors. Voltage application was not necessary during the enzyme reaction, and the response could be preserved for at least 1h.An integrated multi-channel enzyme switch for glucose and lactate was constructed onto the multi-channel electrodes. The integrated enzyme switch showed responses to glucose or lactate independently and the chemical cross-talk was not observed.

酶开关是一种基于覆盖有酶的聚苯胺（PAN）膜的电导测量的新型生物传感器。酶开关可能比常规酶传感器具有以下优点：1）传感芯片本身具有内存函数，2）响应可能独立于电极大小，3）小型化和集成很容易，因为其简单结构。 4）与执行器易于连接，因为电阻变化是指数。在这项研究中，通过使用制备的酶开关来验证这些优势。PAN通过氧化失去其电导率，并通过还原恢复。如果用PAN制备氧化酶 - 过氧化物酶（HRP）共蚀性电极，那么当产生的化合物I和II在PAN表面降低时，PAN可能会被氧化并可能失去电导率。通过监测PAN的电导率，可以通过特定底物的存在来构造“酶 - 开关”，从而在酸性条件下在两个微图案的薄膜电极上进行电化学制备pan膜。然后通过戊二醛交联方法将葡萄糖氧化酶（或乳酸氧化酶）和HRP固定在PAN膜上。通过添加底物，可以观察到由于酶反应而引起的PAN电导率的降低。氧化的锅通过应用0.1 V vs.ag/agcl恢复了其电导率。在流程系统中，大约需要3分钟才能从“ ON”切换到“ OFF”或“ OFF”到“ ON”，并且峰高对应于底物浓度。准备好的酶开关具有酶试管和酶传感器的优点。在酶反应过程中，不需要施加电压，并且至少可以保留1小时的响应。在多通道电极上构建了葡萄糖和乳酸的集成多通道酶开关。综合酶转换显示出对葡萄糖或乳酸的反应，并且未观察到化学交叉对话。