论文总字数：25546字

摘 要

电化学免疫传感器是一种将电化学分析方法与免疫学技术相结合而发展起来的具有快速、灵敏、选择性高、稳定性好，操作简便，检测限低等特点的生物传感器。电化学免疫传感器主要由信号分子，信号扩增分子，电子传递交联复合物，检测物组成。将抗原或者抗体分子固定到电子传递交联复合物表面称为传感器界面的构建，这是免疫传感器的研究和开发中最为关键的步骤。此实验从新型功能电子交联复合物的制备，传感器界面的构建，传感器性能的优化以及免疫传感器的组装等方面进行了探索和研究。此实验创新性的采用了二茂铁与NSA在室温离子液体的环境中组装成FC-NSA电子传递复合物，通过二茂铁独特的电催化氧化还原特性及NSA作为氨基供体可以与多种亲核试剂以酰胺键的形式偶联在一起，同时加入的多壁碳纳米管大大提升整个体系的导电性和结合比表面积，使电流信号提升了1个数量级。为验证实验的成功性，此实验从TEM的组装结构表征，LSV的电聚合表征，免疫传感器的层层组装的循环伏安法表征，以及该传感器的定量分析以及重现性稳定性分析这些方面对本实验设计的免疫传感器效能进行了评测。

关键词：免疫传感器，多壁碳纳米管，N-羟基琥珀酰亚胺丙烯酸酯，二茂铁，离子液体，电化学分析

Abstract

Electrochemical immunosensor is a combination of electrochemical analysis and immunoassay with a host of advantAges. For example, it’s more effective, more sensitive, more stable, more convenient, of lower limit than the other biosensor. Electrochemical immunosensor is constituted by signal element, signal amplifier, electron transferring polymer and analytes. The most critical part of immunosensor is to anchor antibody or antigen to electron transferring polymer. The research makes extensions in several parts, fabrication, optimization, quantitative analysis and so forth. A innovation of the research is adopting Vinylferrocene(FC) and NSA as the electron transferring polymer in the room-temperature ironic liquid(RTIL). The redox activity of FC is shown and NSA, as the provider for amino-group, could bind to plenty of nucleophile due to the formation of amido bond. In the meantime, multi-walls carbon nanotubes(MWCNTs), a conductive material with large specific surface area, is appended to increase the current intensity. To illustrate the results, the research assess the performance of the electrochemical immunosensor by TEM, electro-grafting LSV characterization, CV characterization, quantitative analysis, reproducibility and stability analysis.

KEY WORDS: electrochemical immunosensor, multi-walls carbon nanotubes, NSA, Vinylferrocene, room-temperature ironic liquid, electrochemical analysis

目录

摘要 I

Abstract II

第一章 综述………………………………………………………………………………….6

1.1 免疫测定的手段选择……………………………………………………………6

1.1.1 抗原抗体反应………………………………………………………………6

1.1.2 Sandwich模型……………………………………………………………...6

1.1.3 竞争免疫模型………………………………………………………………6

1.1.4 微流控系统…………………………………………………………………7

1.1.5 等离子共振系统……………………………………………………………7

1.1.6 微悬臂免疫传感器…………………………………………………………8

2.1 免疫测定的识别元件选择……………………………………………………….8

3.1 免疫测定标志物的选择………………………………………………………….9

第二章 引言………………………………………………………………………………..10

第三章 实验部分…………………………………………………………………………..11

1.1 试剂与溶液……………………………………………………………………..11

1.2 仪器……………………………………………………………………………...12

1.3 免疫传感器的制备组装………………………………………………………...12

1.3.1 多壁碳纳米管的制备………………………………………………………12

1.3.2 FC-NSA复合物的制备和表征……………………………………………...12

1.3.3 FC-NSA的电极组装………………………………………………………...13

1.3.4 免疫复合物的组装…………………………………………………………13

1.3.5 免疫复合物的封闭…………………………………………………………13

1.3.5 免疫传感器的抗原修饰……………………………………………………13

第二章 结果与讨论………………………………………………………………………..14

2.1 FC-NSA/MWCNTs的TEM表征……………………………………………………14

2.2 实验条件优化…………………………………………………………………...15

2.2.1 FC-NSA比例的影响………………………………………………………...15

2.2.2 组装MWCNTs的影响………………………………………………………..15

2.2.3 MWCNTs干燥时间的影响…………………………………………………...16

2.3 电聚合的LSV表征……………………………………………………………...17

2.4 电极层层组装的CV表征……………………………………………………….18

2.5 免疫传感器性能测试…………………………………………………………...18

2.5.1 免疫传感器对AFP的定量分析……………………………………………18

2.5.2 免疫传感器的重现性和稳定性……………………………………………19

3.1 结论……………………………………………………………………………...20

致谢……………………………………………………………………………………………21

参考文献………………………………………………………………………………………21

正文

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