基于分子印迹聚离子液体水凝胶阵列的肿瘤标志蛋白电化学传感研究

The objective of this proposal is to prepare molecularly imprinted polymers using poly(ionic liquid) gels and fabricate different nano-array electrochemical sensors with specific recognition ability towards certain template protein (such as tumor markers) in aqueous solution. Based on these new nano-array sensors, electroanalytical methods for the determination of tumor markers with high specificity and sensitivity at low concentration and fast speed under simple conditions can be developed. The new molecularly imprinted nano-array gel is prepared with electrochemical techniques in aqueous solution using an ionic liquid as a functional monomer and a porous polycarbonate film as an array template with a suitable amount of cross-linking agent. The imprinted ratio and mass trasfer rate can be improved based on the control of the structure of nano-array gel, the solubility, the biocompatibility, and the functional groups of ionic liquids which are able to enhance the conformation stability and the interaction between the function monor and the template protein. The weak mechanical strength of poly(ionic liquid) gel is overcomed by using a rigid electrode surface, the sensing surface modified with imprinted polymers is thus able to maintain the most suitable imprinted sites for protein recognition. The relationship between the sensing properties and parameters for the preparation of imprinted materials will be further investigated in order to obtain a better understanding on the mechanism for the recognition of proteins. Moreover, some new techniques for the fabrication of tumor markers imprinted nano-array sensors can be thus developed. Electrochemical behaviors of tumor markers on the molecularly imprinted poly(ionic liquid) gel nano-array surface based on electrochemical probes or signal amplification using nanomaterials will be investigated. The investigations on the signal transmission, the regulation for the enhancement of electrochemical signal, and the sensing properties of the imprinted gels are able to advance the electrochemical sensing methods and techniques for the analysis of trace amount of tumor markers. Real applications are expected for the analysis of tumor makers in real biological samples using this proposed technology.

项目旨在以聚离子液体水凝胶纳米阵列制备分子印迹聚合物，在水溶液中构建对模板蛋白质具有特异识别能力的电化学传感器，建立快速、高选择性、高灵敏度的肿瘤标志蛋白电化学分析新方法。拟采用新型离子液体类功能单体，经适度交联，借助聚碳酸酯阵列模板和电化学调控技术，水相制备对目标蛋白质具有高选择性识别的聚离子液体水凝胶纳米阵列；利用离子液体的水溶性、生物相容性和丰富的、可用于分子识别的官能团，以保持目标蛋白的空间结构的稳定性和多种弱相互作用力，结合纳米阵列结构提高印迹因子和传质速率；依托电极的刚性表面维持水凝胶稳定性，使印迹水凝胶阵列保持最佳的识别位点。系统考察识别特性参数与制备参数之间的内在规律，获取分子印迹阵列传感器的识别机理和制备新原理和新技术；借助电活性探针/纳米信号放大技术，研究肿瘤标志蛋白的信号转换机制、传感特性参数、敏感规律，建立电化学传感新理论及痕量快速检测新方法，并用于实际生物样品分析

项目以肿瘤标志蛋白为模板，离子液体为功能单体，构建分子印迹敏感膜，整合纳米尺寸效应、离子液体优良特性(生物相容性、高导电性、水溶性、可设计性等),构建分子印迹电化学传感器，获取制备原理及技术，探索其识别机制及信号放大规律，并准确分析实际样品中模板蛋白。项目在水相中实现蛋白质印迹过程，有利于保存其空间立体构型及活性，增强印迹因子，提高选择性识别性能，为制备生物大分子印迹敏感膜提供了新思路，新技术及新方法，拓展了离子液体的应用领域。项目完成了以下研究工作：.1、功能化离子液体的制备及表征. 合成含氨基、巯基、吡咯基、乙烯基以及甲基丙烯酸酯等官能团的离子液体，采用核磁、红外光谱、紫外/可见光谱和质谱技术对其进行表征，确保成功制备；.2、离子液体增敏电化学传感性能研究. 基于聚离子液体(或离子液体耦合碳纳米管、石墨烯等)，构建电化学传感界面，研究酚类化合物(如己烷雌酚、双份A、厚朴酚等)电化学行为并检测，证明离子液体的电化学增敏性能；.3、离子液体增敏肿瘤标志蛋白电化学传感. 以离子液体或聚离子液体耦合金纳米颗粒、金纳米棒、根状纳米金、十二面体金纳米晶以及ZnCdHgSe量子点等纳米材料，并负载IgG、前列腺特异性抗原、 神经元特异性烯醇化酶、 鳞状细胞癌抗原等肿瘤标志物的抗体，构建传感平台，研究离子液体在肿瘤标志物电化学传感中的增敏效应及机制，实现其高灵敏、高选择性、高准确度分析；.4、蛋白质印迹聚离子液体电化学传感器. 以牛血清白蛋白、马骨骼肌红蛋白等为模板蛋白，以氨基、乙烯基功能化离子液体为单体，在碳纳米管/玻碳电极界面，室温引发聚合，原位水相制备蛋白质分子印迹敏感膜，研究其分子识别机制及高灵敏测定； .5、甲胎蛋白印迹水凝胶聚离子液体敏感膜及电化学传感. 以甲胎蛋白为模板，乙烯基和甲基丙烯酸酯双功能化离子液体为单体，室温引发聚合，构建分子印迹水凝胶类聚离子液体敏感膜，研究了其水凝胶特征及模板蛋白的选择性识别性能；. 6、阵列式肿瘤标志蛋白分子印迹敏感膜及电化学传感研究. 采用电化学沉积模式，在玻碳电极表面构建金纳米阵列，以双吡咯基功能化离子液体为单体， 神经元特异性烯醇化酶为模板蛋白，电化学聚合，在金纳米阵列间构建分子印迹敏感膜，进而构建阵列式电化学传感体系，实现了神经元特异性烯醇化酶的高选择性、高灵敏度测定。. 研究工作将持续深化。