SPR生物传感器及其与电化学联用研究

发布时间：2018-06-19 11:14

本文选题：表面等离子体共振 + 生物传感器　；参考：《浙江大学》2016年博士论文

【摘要】：随着表面等离子共振(Surface Plasmon Resonance,SPR)生物传感技术的广泛应用,研究SPR技术与其它技术的联用也成为生物传感领域的研究热点。论文首先运用自行研制的SPR生物传感实验仪,开展传感器金属膜系的研究,通过理论分析、仿真计算,得到了金银复合膜最佳厚度设计参数,提高了传感器的性能；结合不同生物识别功能的传感芯片,开展了H7N9亚型流感病毒和环丙沙星(CIP)抗生素的检测实验。在此基础上研究SPR与电化学技术的联用,提出并构建基于二电极体系的ESPR (Electrochemical SPR)系统,结合循环伏安法,开展了甲苯胺蓝(TB)定量检测、聚苯胺(PAn)聚合过程监测等研究。本论文的主要研究内容与成果主要包括：1.研究不同金属膜系对SPR传感性能的影响,通过理论分析与仿真计算分别得到单一金膜、银膜、金银复合膜的最优厚度。实验比较了金银复合膜(40 nm银+10 nm金)与单一金膜(50 nm)的性能。NaCl折射率样品实验,得到金银复合膜的基线噪声为0.004 pixels,折射率分辨率为5.3×10-7 RIU,均优于单一金膜。BSA生物分子吸附实验,结果表明金银复合膜具有更大的SPR共振角响应值。2.运用金银复合膜,结合抗原-抗体识别体系构建了检测H7N9亚型流感病毒的SPR生物传感器。在金属膜表面固定抗H7N9流感病毒HA蛋白的单克隆抗体anti-HA,实现了1～100 ng/mL浓度范围内H7N9病毒的检测,检测限达1ng/mL;anti-HA抗体与HA蛋白的平衡解离常数KD为2.28×10-11 M,表明该anti-HA检测芯片可以有效结合HA蛋白,从而进行H7N9病毒的识别与检测。3.使用金银复合膜,结合分子印迹聚合物(MIP)识别体系,制备了可以特异性检测CIP的MIP芯片。其对CIP具有很好的选择性,检出限为1×10-13mol/L,加标回收率为93.5%～108.8%。同时,该检测芯片可再生并重复利用。4.研究SPR与电化学联用技术,提出以活性炭为辅助电极的二电极体系,构建了二电极体系ESPR生物传感器。采用循环伏安法进行不同浓度TB溶液的检测实验,根据电流响应曲线可得到TB发生氧化还原反应的氧化峰、还原峰电位信息。将SPR共振角数据对电位求导得到E-dθ/dt曲线,其变化与法拉第电流相同,在谱图上同样出现谱峰,且峰电位与循环伏安法得到氧化峰、还原峰电位一致。将不同浓度TB溶液的氧化峰峰位电流值和dθ/dt数值分别与TB溶液浓度进行拟合,线性相关性较好,可进行TB溶液的浓度检测。5.使用二电极体系ESPR系统,结合循环伏安法,实时监测苯胺单体在SPR传感金属膜表面合成PAn的过程。循环伏安电流曲线只能得到简单的氧化还原峰电位信息,而通过SPR共振角曲线可以了解到PAn在不同氧化还原态之间的相互转化过程,进而分析PAn的增长过程动力学,为研究PAn的聚合机理提供新方法。
[Abstract]:With the wide application of Surface Plasmon Resonance (SPR) biosensor technology, the research on the combination of SPR and other technologies has become a hotspot in the field of biosensor. Firstly, the SPR biosensor is used to study the metal film system of the sensor. Through theoretical analysis and simulation calculation, the optimum thickness design parameters of the gold and silver composite film are obtained, and the performance of the sensor is improved. The detection of H7N9 subtype influenza virus and ciprofloxacin (CIP) antibiotics was carried out with different biometric sensing chips. On this basis, the combination of SPR and electrochemical technology was studied, and the ESPR / Electrochemical SPRs system based on two-electrode system was proposed and constructed. Combined with cyclic voltammetry, the quantitative detection of toluidine blue (TB) and the monitoring of Polyaniline (Polyaniline) Polyaniline (pan) polymerization process were carried out. The main contents and achievements of this thesis include: 1. The effects of different metal film systems on the sensing performance of SPR were studied. The optimal thickness of single gold film, silver film and gold-silver composite film was obtained by theoretical analysis and simulation. The properties of gold and silver composite films (40 nm silver 10 nm gold) and a single gold film (50 nm) were compared with those of NaCl refractive index samples. The base-line noise and refractive index resolution of gold-silver composite film are 0.004 pixels and 5.3 脳 10 ~ (-7) RIUrespectively, which are superior to the single gold film. BSA biomolecular adsorption experiment. The results show that the gold and silver composite film has a larger resonance angle response value of .2. A SPR biosensor for detecting H7N9 subtype influenza virus was constructed by using gold and silver composite membrane and antigen-antibody recognition system. The monoclonal antibody anti-HAA against HA protein of H7N9 influenza virus was immobilized on the surface of metal membrane, and the detection of H7N9 virus was achieved in the concentration range of 100 ng / mL of H7N9 virus. The equilibrium dissociation constant of anti-HA antibody and HA protein was 2.28 脳 10 ~ (-11) M, which indicated that the anti-HA detection chip could effectively bind HA protein and detect H7N9 virus. Using gold and silver composite membrane and molecular imprinted polymer (MIP) recognition system, a MIP chip for specific detection of CIP was prepared. It has good selectivity to CIP, the detection limit is 1 脳 10 ~ (-13) mol / L, and the recovery rate is 93.5% ~ 108.8%. At the same time, the chip can be regenerated and reused. A two-electrode system with activated carbon as auxiliary electrode was proposed and ESPR biosensor was constructed. The cyclic voltammetry was used to detect the TB solution with different concentrations. According to the current response curve, the redox peak and the reduction peak potential of TB were obtained. The E-d 胃 / DT curve is derived from the SPR resonance angle data. The variation is the same as that of Faraday current, and the peak value is the same as that of Faraday current, and the peak potential is the same as the oxidation peak and the reduction peak potential obtained by cyclic voltammetry. The oxidation peak current value and d 胃 / DT value of TB solution with different concentrations were fitted to the concentration of TB solution respectively. The linear correlation was good, and the concentration of TB solution could be detected by .5. Using the two-electrode ESPR system and cyclic voltammetry, the synthesis of pan on the surface of SPR sensing metal membrane by aniline monomer was monitored in real time. The cyclic voltammogram current curve can only obtain the simple information of the redox peak potential, but the SPR resonance angle curve can be used to understand the interconversion process of pan in different redox states, and then analyze the kinetics of pan growth process. It provides a new method for studying the polymerization mechanism of pan.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：O657.1;TP212.3

【相似文献】