复合材料组装的免疫传感器用于农药克百威残留检测研究

毕业论文

复合材料组装的免疫传感器用于

农药克百威残留检测研究

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年 月摘 要

I 摘 要

本实验制备了一种新型的电化学免疫传感器，以纳米金、普鲁士蓝/多壁碳/壳聚糖（PB/MWCNTs/CHIT）复合膜和蛋白A为基础对克百威农药残留进行检测。纳米金增加电极表面电子传导能力和稳定性，PB/ MWCNTs/CHIT复合膜提供了更多的氨基和羧基，能够更好的在电极上交联组装蛋白A增加定向固定抗体的能力。用循环伏安法（CV）、差分脉冲伏安法(DPV)、方波伏安法(SWV)描述免疫传感器的组装过程的特性。并对实验参数进行优化，如纳米金的电沉积时间、测试底液的pH、抗原的孵育时间。在最适条件下，电流变化量与克百威的浓度对数成线性关系，在1ng/ml到1μg/mL和1μg/mL到200μg/mL呈良好的线性关系，其相关系数分别为0.9951和0.9963，检出限为0.533 ng/mL。本免疫传感器具有较高的灵敏度、良好的稳定性、能被重复使用，可以用来对克百威杀虫剂残留的检测。

关键词：免疫传感器，纳米金，PB/MWCNTs/CHIT复合膜，蛋白A，克百威 ABSTRACT

II ABSTRACT

In this paper, a novel electrochemical immunosensor for the determination of

carbofuran based on nanogold particles (nano-Au), prussian blue/multi-walled carbon

nanotubes/chitosan (PB/MWCNTs/CHIT) and protein A (SPA) composite film was

proposed.Nano-Au,PB and MWCNTs-incorporated film was used to enhance the

electroactivity and stability of the immunosensor. PB/MWCNTs/CHIT composite film

provided many amino groups and carboxyl groups to cross-link SPA.We have

employed the formation of aself-assembled proteinA layer on a electrode to increase

binding capacity to antibody. The conformational properties of the immunosensor and

electrochemical performance of immunoreaction between antigen and the

anti-carbofuran monoclonal antibody (Ab) were characterized using Difference pulse

volt-ampere(DPV), cyclic voltammetry (CV) and square wave voltammetry (SWV).

Effects of experimental variables, such as the adsorption time of gold nanoparticles,

the pH of supporting electrolyte,and the incubation time were investigated in

details.Under optimal conditions, the current response was proportional to the

concentration of carbofuran which ranged from 1ng/ml to1μg/mL and 1μg/mL to

200μg/mL with correlation coefficients of 0.9951 and 0.9963, respectively.The

detection limit was 0.533ng/mL.It was concluded that the proposed immunosensor

exhibited high sensitivity, high stability and good reproducibility, and it can be used

for detection of carbofuran pesticide.

Keywords: immunosensor, nanogold particles, PB/MWCNTs/CHIT, protein A,

carbofuran 目 录

III 目 录

摘 要 .............................................................................................................. I

ABSTRACT ..................................................................................................... II

目 录 ............................................................................................................ III

第一章 绪 论 .............................................................................................. 1

1.1 引言 ............................................................................................................ 1

1.2 生物传感器简介 .......................................................................................... 1

1.2.1 生物传感器的原理 ............................................................................. 1

1.2.2 生物传感器的分类 ............................................................................. 2

1.2.2.1 酶传感器 ...................................................................................... 2

1.2.2.2免疫传感器 ................................................................................... 3

1.2.2.3 DNA传感器 .................................................................................. 4

1.2.2.4 微生物传感器 .............................................................................. 4

1.2.2.5 其他类型生物传感器 ................................................................... 5

1.2.3 生物传感器的应用 ............................................................................. 6

1.2.3.1 在医学领域中的应用 ................................................................... 6

1.2.3.2 在食品安全上的应用 ................................................................... 6

1.2.3.3 在环境监测中的应用 ................................................................... 6

1.3 本课题研究的内容、存在问题及发展趋势 ................................................... 7

1.3.1 存在的问题及发展趋势 ..................................................................... 7

1.3.2 研究目标 ............................................................................................ 8

1.3.3 本课题研究内容 ................................................................................. 8

第二章 实验材料与方法 ................................................................................. 9

2.1 实验材料 ..................................................................................................... 9

2.1.1 主要试剂 ............................................................................................ 9

2.1.2 主要仪器 .......................................................................................... 10