基于二维有序球腔阵列微电极的蛋白质电化学传感器研究

项目名称： 基于二维有序球腔阵列微电极的蛋白质电化学传感器研究

项目编号： No.21505079

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 顾学芳

作者单位： 南通大学

项目金额： 21万元

中文摘要： 为实现对癌症标志蛋白的超高灵敏检测，本项目拟通过构筑一种具有二维有序球腔结构的微电极阵列，结合杂交链式反应技术及免疫识别形成“超级三明治”式的免疫复合物，以电化学手段为信号读出方法来加以解决。首先，利用自组装在电极表面制备二维有序的聚苯乙烯模板，进而通过电化学沉积结合牺牲模板法构筑系列底部开放的二维有序绝缘体（如SiO2）及半导体（如WO3）微腔阵列，以形成电极表面电化学微反应区。其次，通过杂交链式反应在引入带有标记分子的DNA链节连环，构建实现标记分子电化学信号的原位扩增，提高检测灵敏度。本课题拟在解决新型蛋白质电化学传感器可控制备的基础上，阐明此类球腔微电极阵列的的微结构控制机制及电化学特性，建立电化学检测模型；通过微球腔内表面的修饰，探寻有利于保持蛋白分子活性及电子传递的微环境体系；揭示标记分子扩增体系及抗体在金纳米颗粒表面的竞争吸附行为与信号灵敏度及重现性之间的内在联系等基础问题。

中文关键词： 微电极阵列；杂交链式反应；电化学免疫分析；信号增强

英文摘要： Cancer is a serious disease that jeopardizes human life and health, however, a fundamental breakthrough in human cancer treatment is still missing. Early detection and early treatment are still the basic countermeasures to treat cancer patients. Therefore, to achieve the ultra-sensitive detection of cancer biomarkers, in this project, an electrochemical-based protein sensor with a novel “super-sandwich” structure, which consists of two-dimensional highly-ordered nanocavity array and signal amplified immunocomplex on gold nanoparticles, will be designed and synthesized for early diagnosis of cancer biomarkers. Firstly, a two-dimensional ordered polystyrene template will be prepared on the surface of ITO electrodes through self-assembly technique, combined with electrochemical deposition and a sacrificial-template method, series of two-dimensional ordered insulators (such as SiO2) and semiconductors (such as WO3) microcavity array will be constructured, serving as the electrochemical microreactor on the the electrode surface. Secondly, a long DNA concatemer with redox-active probe molecules will be desiged using hybridization chain reaction (HCR) technique, which accordingly achieves significant in situ signal amplification to enhance the detection sensitivity. Using several typical cancer protein markers as research models, through control group tests，the diagnosis value of the proposed protein chip will be evaluated. Besides the controllable preparation of the novel electrochemical sensor for cancer biomarker detection, the microstructure-control mechanism and the electrochemical properties of such microcavity array electrodes will be clarified; the model for electrochemical detection on such electrodes will also be established. Moreover, the suitable micro-environment for keeping protein activity and for electrons transfer will be investigated through the modification of the inner surface of the as-prepared microcavity array. Additionally, the internal connection between signal sensitivity and the competitive adsorption behavior (occurred between the antibody and the labeled–molecule-amplification system) will be revealed.

英文关键词： microelectrode array;hybridization chain reaction;electrochemical immunoassay;signal amplification