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Development of Sol-Gel Silicate Based Mictoarray Biosensors

基于溶胶-凝胶硅酸盐的微阵列生物传感器的开发

基本信息

批准号：
09640732
负责人：
YAO Toshio
金额：
$ 1.73万
依托单位：
Osaka Prefecture University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

The sol-gel method has been demostrated as a low-temperature technique for the production of new inorganic materials. In this work, sol-gel silicate-based biosensors were made by utilizing a composite membrane of sol-gel enzyme film and electrochemically generated poly(1 , 2-diaminobenzene) film to improve the selectivity of the sol-gel enzyme sensors. However, the prepared sensor was poor in operational stability, because of the release of enzyme entrapped into the sol-gel matrics. Therefore, to prevent the leaching of entrapped enzyme, the prepared sol-gel enzyme sensor was exposed to glutaraldehyde vapor. This treatment was effective to improve the stability of the sensor, but tended to decrease the sensitivity because of enzymic inactivation. To improve these problems of sol-gel silicate based biosensor, the biotin-avidin affinity reaction was adopted for the production of the enzyme receptor composed of sol-gel matrics. Avidin, having four biotin binding sites, crosslinked and immobilized enzyme labeled with multiple biotin into the sol-gel silicate lattice, when tetramethyl orthosilicate solutions of biotin labeled enzyme and avidin were mixed on the electrode. The prepared sol-gel sensors were excellent in both stability and sensitivioty. This preparation method was applied to the construction of multiarray biosensors with detection units of glucose, cholesterol, total cholesterol, and urate, and of orthophosphate, total orthophosphate, and ammonia, and the construction of freshness sensors of fish and meat.

溶胶-凝胶法是一种低温制备新型无机材料的方法。本工作利用溶胶-凝胶酶膜和电化学生成的聚对苯二胺膜的复合膜制备了溶胶-凝胶硅酸盐基生物传感器，以提高溶胶-凝胶酶传感器的选择性。然而，所制备的传感器的操作稳定性差，因为在溶胶-凝胶基质中捕获的酶的释放。因此，为了防止包埋酶的浸出，制备的溶胶-凝胶酶传感器暴露于戊二醛蒸汽。这种处理有效地提高了传感器的稳定性，但由于酶的失活而倾向于降低灵敏度。针对溶胶-凝胶硅酸盐生物传感器存在的这些问题，采用生物素亲和素亲和反应制备溶胶-凝胶基质酶受体。当生物素标记的酶与亲和素的正硅酸四甲酯溶液混合在电极上时，具有四个生物素结合位点的亲和素将多个生物素标记的酶交联并固定到溶胶-凝胶硅酸盐晶格中。所制备的溶胶-凝胶传感器具有良好的稳定性和灵敏度。这种制备方法被应用于构建多阵列生物传感器与检测单元的葡萄糖，胆固醇，总胆固醇，尿酸盐，正磷酸盐，总正磷酸盐，和氨，和建设新鲜度传感器的鱼和肉。