功能化纳米复合材料构建的免疫传感器用于肿瘤标志物检测

发布时间：2018-01-07 16:04

本文关键词：功能化纳米复合材料构建的免疫传感器用于肿瘤标志物检测　出处：《湖北民族学院》2017年硕士论文　论文类型：学位论文

【摘要】：金属纳米材料(金纳米粒子、银纳米粒子、铂纳米粒子)和碳纳米材料(氧化石墨烯,碳纳米管)具有超强的导电性,良好的生物相容性以及巨大的比表面积等优良的物理化学性能,被广泛的应用于免疫传感器中。将这些纳米材料经功能化复合后形成的功能化纳米复合材料,其各项性能都大大的增强。基于此本论文构建了肿瘤标志物检测的新方法,其主要内容包括以下五章:第一章:绪论。简要概述癌症的发病机理及其检测方法,免疫传感器的分类及检测原理,详细介绍金属纳米材料和碳纳米材料在免疫传感器的应用,最后介绍本论文的研究内容。第二章:实验基于纳米铂-单壁碳纳米管-壳聚糖纳米复合材料修饰玻碳电极,并通过纳米铂及富含羧基的单壁碳纳米管将癌胚抗原抗体固定到电极表面制成了免疫传感界面。选用癌胚抗原(CEA)作为分析模型,构建一种简单灵敏无标记的阻抗型免疫分析方法。在最优化的条件下,CEA浓度在0.01~12 ng/m L范围时,阻抗值随着CEA的浓度增加而增加并表现出良好的线性关系,检测限为0.0042 ng/mL(S/N=3)。该免疫传感器具有良好的稳定性和选择性,回收率在94.6%~108.6%。第三章:实验以性能优良的十二面体金-银核壳型纳米晶体复合材料与前列腺特异性抗体结合制备免疫复合物为信号探针,金电极上电沉积一层银纳米花后与金纳米/氧化石墨烯/壳聚糖混合碳晶胶纳米材料复合制得的免疫传感平台,采用夹心型免疫分析模式,建立了信号抑制型的前列腺特异性抗原免疫传感器。其中银纳米花(AgNFs)作为氧化还原电子媒介体,并随着PSA浓度的增加,免疫传感器上形成的免疫复合物越多越多抑制银的电化学响应,导致了DPV峰电流信号逐渐减小。在最优化条件下,免疫传感器对PSA检测的范围为1.0×10-5~20.0 ng/mL,检测限可达到3.6×10-6 ng/mL(S/N=3)。第四章:实验采用功能化修饰的金纳米棒捕获甲胎蛋白抗体形成纳米复合物,通过电沉积在金电极上的银纳米花将纳米复合物修饰到电极表面,制得电化学免疫传感器,构建了一个简单灵敏无信号标记的甲胎蛋白(AFP)检测新方法。检测范围为0.001~20 ng/m L,检测限为0.5 pg/m L(S/N=3),对临床样品的分析结果与商用电化学发光法一致。第五章:实验将制备的爆米花状的金纳米粒子作为分子标记物同时捕获甲胎蛋白抗体和辣根过氧化酶形成信标探针。同时以氧化石墨烯-亚甲基蓝-金纳米粒子纳米复合材料滴涂到玻碳电极表面上固定甲胎蛋白抗体构建免疫传感平台。采用夹心型的免疫分析模式,建立了一种新型电化学免疫传感分析方法用于检测甲胎蛋白(AFP)。在优化的实验条件下,该免疫传感器对AFP检测限为0.32 pg/m L(S/N=3),并能用于实际样品的检测中。
[Abstract]:Metal nanoparticles (gold nanoparticles, silver nanoparticles, platinum nanoparticles) and carbon nanomaterials (graphene oxide, carbon nanotubes) have super conductivity. Excellent physical and chemical properties such as good biocompatibility and large specific surface area are widely used in immunosensors. Based on this paper, a new method for detection of tumor markers has been constructed. The main contents include the following five chapters: chapter 1: introduction. A brief overview of the pathogenesis of cancer and its detection methods. The classification and detection principle of immunosensor, the application of metal nano-material and carbon nano-material in immunosensor are introduced in detail. Finally, the research content of this thesis is introduced. Chapter 2: the experiment is based on the modified glassy carbon electrode based on nano-platinum single-walled carbon nanotube-chitosan nanocomposites. The immunosensor interface was made by immobilization of carcinoembryonic antigen antibody to the electrode surface by platinum nanoparticles and carboxyl-rich single-walled carbon nanotubes. Carcinoembryonic antigen (CEA) was used as the analytical model. A simple, sensitive and unlabeled impedance immunoassay was constructed. Under the optimal conditions, the concentration of CEA was in the range of 0.01 ~ 12 ng/m / L. The impedance value increases with the concentration of CEA and shows a good linear relationship. The detection limit is 0.0042 ng / mL / S / N ~ (3 +). The immunosensor has good stability and selectivity. The recovery rate is 94.6 and 108.6. Chapter III:. In the experiment, the immune complex was prepared by combining 12 hedron gold-silver core-shell nanocrystalline composite with prostate specific antibody as signal probe. The immunosensor platform was prepared by electrodepositing a layer of silver nanosplast and mixed with gold nanocrystalline / graphene oxide / chitosan on the gold electrode. The sandwich immunoassay model was used. A signal suppressive prostate specific antigen immunosensor was established in which AgNFs was used as redox electronic medium and the concentration of Agns increased with the increase of PSA concentration. The more immune complexes formed on the immunosensor, the more inhibition of the electrochemical response of silver, which led to the gradual decrease of DPV peak current signal. The detection range of PSA by immunosensor was 1.0 脳 10 ~ (-5) ng/mL. The detection limit can reach 3.6 脳 10 ~ (-6) ng 路m ~ (-1) mg / m ~ (-1) S / N ~ (3). Chapter 4th: functional modified gold nanorods were used to capture alpha-fetoprotein antibodies to form nanocomplexes. An electrochemical immunosensor was prepared by modifying the nano-composite onto the surface of the electrode by electrodeposition of silver nanoparticles on the gold electrode. A simple and sensitive AFP method for detection of alpha-fetoprotein was constructed with a detection range of 0.001 ~ 20 ng/m / L. The detection limit was 0.5 pg/m L ~ (-1) S / N ~ (3). The results of analysis of clinical samples are consistent with those of commercial electrochemiluminescence. Chapter 5th:. The prepared popcorn gold nanoparticles were used as molecular markers to capture alpha-fetoprotein antibody and horseradish peroxidase simultaneously to form a beacon probe. The material was coated on the surface of glassy carbon electrode to immobilize AFP antibody to construct the immunosensor platform. The sandwich immunoassay model was used. A novel electrochemical immunosensor method was developed for the detection of AFPN. Under the optimized experimental conditions, a novel electrochemical immunosensor method was developed for the detection of AFP. The detection limit of the immunosensor for AFP is 0.32 pg/m / L ~ (-1) S / N ~ (3 +), and it can be used in the detection of real samples.
【学位授予单位】：湖北民族学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R730.4;O657.1

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