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一种有前途的新型生物分子固定基质:含功能苯并咪唑的导电聚合物的合成及其生物传感器应用。

A novel promising biomolecule immobilization matrix: synthesis of functional benzimidazole containing conducting polymer and its biosensor applications.

机构信息

Department of Polymer Science and Technology, Middle East Technical University, 06800 Ankara, Turkey.

出版信息

Colloids Surf B Biointerfaces. 2013 Dec 1;112:74-80. doi: 10.1016/j.colsurfb.2013.07.049. Epub 2013 Aug 2.

Abstract

In order to construct a robust covalent binding between biomolecule and immobilization platform in biosensor preparation, a novel functional monomer 4-(4,7-di(thiophen-2-yl)-1H-benzo[d]imidazol-2-yl)benzaldehyde (BIBA) was designed and successfully synthesized. After electropolymerization of this monomer, electrochemical and spectroelectrochemical properties were investigated in detail. To fabricate the desired biosensor, glucose oxidase (GOx) was immobilized as a model enzyme on the polymer coated graphite electrode with the help of glutaraldehyde (GA). During the immobilization step, an imine bond was formed between the free amino groups of enzyme and aldehyde group of polymer. The surface characterization and morphology were investigated to confirm bioconjugation by X-ray photoelectron spectroscopy (XPS) and transmission electron microscopy (TEM) at each step of biosensor fabrication. The optimized biosensor shows good linearity between 0.02mM and 1.20mM and a low limit of detection (LOD) of 2.29μM. Kinetic parameters Km(app) and Imax were determined as 0.94mM and 10.91μA, respectively. The biosensor was tested for human blood serum samples.

摘要

为了在生物传感器制备中构建生物分子与固定化平台之间的牢固共价键,设计并成功合成了一种新型功能单体 4-(4,7-二(噻吩-2-基)-1H-苯并[d]咪唑-2-基)苯甲醛(BIBA)。该单体聚合后,详细研究了其电化学和光谱电化学性质。为了制备所需的生物传感器,葡萄糖氧化酶(GOx)在戊二醛(GA)的帮助下作为模型酶固定在聚合物涂层石墨电极上。在固定化步骤中,酶的游离氨基与聚合物的醛基之间形成亚胺键。通过 X 射线光电子能谱(XPS)和透射电子显微镜(TEM)在生物传感器制备的每个步骤对表面特征和形态进行了研究,以确认生物结合。优化后的生物传感器在 0.02mM 至 1.20mM 之间呈现出良好的线性关系,检测限(LOD)低至 2.29μM。动力学参数 Km(app)和 Imax 分别为 0.94mM 和 10.91μA。该生物传感器已用于人血清样品的测试。

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