基于仿生离子通道识别的电位型传感器检测海水中浮游病毒

Research on sensor technology for monitoring of virioplankton in seawater will provide important technical support for efficient marine eco-environmental monitoring and early warning. Also, it can satisfy the urgent need of our country in marine monitoring. In this project, potentiometric sensors based on biomimetic ion channel for the detection of virioplankton in the presence of a high interfering background will be systematically investigated. The sensing mechanism of the proposed senors in separating and enriching virioplankton and responding to the indicator ion in seawater will further be discussed. The ion channel is prepared by choosing suitable preparation conditions. In order to improve the selectivity of the obtanined ion channel and eliminate the matrix effects and complicated interference from seawater, molecularly imprinted polymer (MIP) which is systhesized under optimized polymerization conditions is employed to functionlize the channel surface. The indicator ions are driven by the applied current pulse to diffuse from the solution phase into the ion channel and finally into the sensing membrane. The recognition interactions between virioplankton and the MIP in the channel surface are indicated by the current-driven ion fluxes. Thus, virioplankton can be indirectedly detected by the potentiometric sensor based on the MIP functionlized ion channel. By optimizing the chronopotentiometric detection conditions, the electrode sensitivity can be largely improved. For the first time, a novel potentiometric sensor for the detection of marine virioplankton is developed. This study makes a great contribution to the improvement of sensor sensitivity and selectivity and analysis efficiency on the determination of virioplankton in seawater, which will significantly promote the development of sensor technology in the field of marine monitoring.

海水中浮游病毒检测传感器技术的研究可为及时高效地开展海洋生态环境监测及预警提供重要的技术手段，为满足国家在海洋监测领域的重大需求提供科技支撑。本项目对基于仿生离子通道的电位型传感器在强电解质干扰条件下检测病毒进行系统研究，深入探讨在海水基体条件下传感器对浮游病毒的分离富集原理及对指示离子的响应机理。通过筛选离子通道的制备条件，优化聚合反应条件，改善分子印迹聚合物修饰的离子通道孔道对病毒分子识别的特异性，消除海水基体效应及复杂成分的干扰，实现浮游病毒的高选择性分离富集；采用电流法驱动指示离子通过离子通道进入电极敏感膜中，以指示病毒与离子通道内印迹聚合物之间的选择性键合作用；通过优化计时电位检测条件，提高电极检测灵敏度，首次开发出一类检测海水中浮游病毒的新型电位型传感器，在提高对海水中病毒高选择性识别和快速、高灵敏检测等方面具有重要的理论和方法创新，为满足国家在海洋监测领域的需求做出贡献。

发展适合于现场快速检测海洋浮游病毒的传感器技术，对于及时快速地开展海洋环境监测和预警具有重要意义。目前，海水中浮游病毒的所有定性定量分析方法均仅适用于实验室分析，不能进行现场快速监测。聚合物膜离子选择性电极适于现场监测，是海水浮游病毒检测的理想传感器。然而，此类电极适于检测有机或无机小离子，而不适于高电荷大体积病毒分子的检测。近年来，离子通道电位型传感器技术的出现，有望实现病毒的电位检测。但是，离子通道的选择性较差，这严重影响了传感器响应性能。.本研究利用分子印迹聚合物修饰离子通道，提高离子通道识别能力；将仿生离子通道覆盖于敏感膜表面，采用电流驱动指示离子指示选择性识别作用，利用计时电位法提高检测速度和灵敏度，发展基于仿生离子通道的电位型传感器。.项目合成了分子印迹聚合物修饰的离子通道，考察了其识别性能；构建了基于计时电位法的仿生离子通道电位型传感器，优化了计时电位检测条件；利用电流驱动指示离子进出电极敏感膜，探讨了电位法快速检测浮游病毒分子的新机理，实现了浮游病毒的高灵敏、高选择性电位检测，电极对100-5000 copy/mL的对虾白斑综合症病毒呈现线性响应，检出限可达50 copy/mL；制备了仿离子通道电位型传感器，拓宽了离子通道电位传感技术应用范围；发展了诸多选择性好、可逆性高、长期稳定性优越的电位型传感器，为浮游病毒电位型传感器在海洋监测中的应用提供技术支持。 .项目开发的分子印迹离子通道技术，将显著提升基于离子通道技术的光、电化学传感器响应性能；开发的电位法检测蛋白质、病毒、细胞的方法，将极大地拓宽电位分析技术的应用空间。.共SCI 期刊论文14 篇，其中1篇发表于《德国应用化学》、3篇发表于《美国分析化学》；培养博士及硕士毕业生4名；在国内外会议上做邀请/口头报告6次；研究团队获得2017年度海洋工程科技奖一等奖以及2018年度中国分析测试协会科学技术奖一等奖。