CHANGSHA UNIVERSITY OF SCIENCE & TECHNOLOGY
毕业设计（论文）
题目：基于纳米自组装单分子膜的QCM传感分析研究
学生姓名： 张元元 学 号： 200729230210 班 级: 环工0702 专 业： 环境工程 指导教师： 曹 忠
2011 年 6 月
基于纳米自组装单分子膜的QCM传感分析研究
学生姓名： 张元元 学 号： 200729230210 班 级： 07-02
所在院(系): 化学与生物工程学院 指导教师： 曹 忠 完成日期: 2011年6月
基于纳米自组装单分子膜的QCM传感分析研究
基于纳米自组装单分子膜的QCM传感分析研究
摘要
本论文的实验是用L-半胱氨酸修饰电极表面，戊二醛共价交联法包被抗体，最后用纳米金标记二抗进行放大的方法检测蛋白，对压电免疫传感器进行了探索研究。首先在压电石英晶体金电极表面自组装一层带巯基的半胱氨酸单分子膜，再在膜上组装一层戊二醛，通过吸附作用，将c-myc抗体固定于石英晶体表面。通过c-myc抗体与c-myc蛋白的特异性反应将c-myc蛋白固定于晶振表面，最后用纳米金标记的二抗GAM放大抗原-抗体的识别信号。用QCM对纳米金标记的二抗和一抗识别的蛋白进行连续检测，实验结果表明L-半胱氨酸固定前后频率的大致变化范围为800Hz-1800Hz，戊二醛修饰前后频率的变化范围为1200Hz-3300Hz，c-myc抗体固定前后频率变化范围为1000Hz-3000Hz，一抗与抗原发生特异性结合前后频率变化的大致范围为1000Hz-2800Hz，c-myc蛋白浓度在8.61×10-12 mol/L～1.13×10-10 mol/L之间，随着c-myc蛋白浓度的增加频率的变化量增大，二抗GAM成功固定。传感器经再生，频移值约在士100Hz范围内变动，传感器容易再生。
关键词：纳米金；自组装单分子膜；QCM；c-myc
基于纳米自组装单分子膜的QCM传感分析研究
BASED ON NANO SELF-ASSEMBLED MONOLAYERS OF
QCM SENSING ANALYSIS
ABSTRACT
Experiments of this paper is to use L-cysteine modified electrode surface, glutaraldehyde covalent crosslinking coating antibody, and finally labeled with gold nanoparticles to amplify the secondary antibody used to detect the protein on the piezoelectric immunosensor was explored study. First, the surface of piezoelectric quartz crystal gold electrode with thiol self-assembled layer of the cysteine monolayer, then assembled in the membrane layer of glutaraldehyde, through adsorption, the c-myc antibody fixed on the quartz crystal surface. By c-myc and c-myc protein antibody specific to c-myc protein in response to fixed on the crystal surface, finally, gold nanoparticles labeled secondary antibody GAM can significantly enlarge the antigen - antibody recognition signal. QCM on gold nanoparticles labeled with secondary antibody and a protein recognized by anti-continuous testing. The results show that L-cysteine or less constant before and after the change in the frequency range of 800Hz-1800Hz, Glutaraldehyde modification of the range before and after the change of frequency 1200Hz-3300Hz, c-myc antibody before and after a fixed frequency range 1000Hz-3000Hz, An anti-specific binding of antigen with the approximate frequency range before and after the 1000Hz-2800Hz, In 8.61 × 10-12 mol / L ~ 1.13 × 10-10 mol / L, between c-myc protein with increasing concentrations of the change in frequency increases, Second, successful anti-GAM fixed. The regeneration of the sensor frequency shift value of about ± 100Hz range of changes in the sensor easy to regenerate.
Key words: Gold nanoparticles; Self-assembled monolayers; QCM; c-myc
基于纳米自组装单分子膜的QCM传感分析研究
目 录
1 绪论.....................................................................................................................................1 1.1 QCM概述....................................................................................................................1 1.2 纳米金自组装膜的简介和研究进展..........................................................................2 1.2.1 自组装单分子膜.................................................................................................2 1.2.2 纳米金概述.........................................................................................................3 1.2.3 纳米金标记技术.................................................................................................4 1.2.4 纳米金自组装单分子膜的研究进展.................................................................5 1.3 基于纳米自组装膜的QCM应用...............................................................................6 1.3.1纳米颗粒用于提高生物传感器的灵敏度...........................................................6 1.3.2纳米磁颗粒用于提高再生能力和缩短检测时间...............................................7 1.4 C-myc蛋白..................................................................................................................8 1.5 本文的研究内容..........................................................................................................8 2 实验部分..............................................................................................................................9 2.1 实验仪器与试剂...........................................................................................................9 2.1.1 仪器......................................................................................................................9 2.1.2 实验试剂..............................................................................................................9 2.1.3 溶液配制.............................................................................................................10 2.2 实验过程......................................................................................................................11 2.2.1 纳米金的制备.....................................................................................................11 2.2.2 L-半胱氨酸的固定.............................................................................................12 2.2.3戊二醛共价交联法抗体包被..............................................................................12 2.2.4 c-myc蛋白的固定..............................................................................................12 2.2.5合成纳米金的标记..............................................................................................12 2.2.6二抗的固定..........................................................................................................12
2.2.7传感器的再生.....................................................................................................12
2.2.8 晶振修饰前后的表征........................................................................................12
3 结果与讨论.........................................................................................................................14