基于导电聚合物构建的甲胎蛋白免疫传感器的制备和研究

发布时间：2018-04-12 07:01

本文选题：甲胎蛋白 + 导电聚合物　；参考：《石河子大学》2016年硕士论文

【摘要】：研究表明,目前唯一可以用来筛选和检测肿瘤标志物的重要标志是人体血清中甲胎蛋白(AFP)的含量,及时发现与诊断肿瘤的出现可以准确且灵敏地检测出血清中的AFP含量。传统免疫监测方法由于分析时间长、分析仪器昂贵及需要专业操作人员等限制了其发展,此时具有操作简单、灵敏度高、测定时间较短及成本低等特点的免疫传感器得到了研究者的强烈关注。本文根据不同的电极修饰材料制备了高灵敏度及工作性能稳定的电化学免疫传感器。电化学免疫传感器中我们主要研究的是敏感界面电活性材料的修饰。在本文中我们应用的是不易在电极表面流失,易于固定在电极表面的导电高分子聚合物。导电聚合物目前在传感器中广泛应用主要是由于其优良的性质(环境友好性、高电导率、稳定性好等)。导电聚合物如聚苯胺(PANI)、聚吡咯(PPy)、聚邻苯二胺(PoPD)等经过掺杂后导电性能有着很大的改善,利用此类型材料构建的电化学免疫传感器的性能能够得到很大的提升。最近几年,仿生纳米材料因其具有成膜性优良、制备简单及生物相容性好的特性,在免疫传感器的研制中成为研究方向的着力点之一。聚多巴胺目前作为一类很好的二级反应平台引起了人们的广泛关注,这主要是由于聚多巴胺的结构中含有氨基和酚羟基,从而使其生物相容性良好,是目前众多生物友好型大分子中的一种。本论文采用电聚合及电沉积方法将导电聚合物固定在电极上从而构建出一系列高度灵敏和稳定性优良的电化学免疫传感器;试验中采用电化学分析技术对免疫电极的自组装过程和电极界面进行了表征并探讨了免疫电极在生物分析方面的应用。本论文主要研究内容如下:1.基于掺杂态PANI构建的甲胎蛋白免疫传感器:组装完成了具有有高选择性、稳定性、灵敏性的新型非标记型电流型免疫传感器,主要用于甲胎蛋白(AFP)的检测。采用电聚合方法将聚苯胺薄膜修饰在玻碳电极表面,接着将甲胎蛋白抗体(anti-AFP)修饰在聚苯胺表面。实验表明此免疫传感器的线性范围为0.01-50 ng/mL,检出限可以达到0.008 ng/mL。此免疫传感器在制备过程中体现出了选择性高、重现性及稳定性好的优点。2.基于PPy及PDA构建的甲胎蛋白免疫传感器:聚吡咯/聚多巴胺(PPy/PDA)非标记型电化学免疫传感器的成功组装,可以用于检测甲胎蛋白(AFP)。采用电沉积方法将吡咯(Py)沉积在玻碳电极表面形成聚吡咯(PPy)膜,然后利用多巴胺(DA)在弱碱性溶液中可以自氧化聚合,在PPy表面包覆成膜修饰在电极表面以固定anti-AFP。聚多巴胺膜的优良的生物相容性为该传感器固定抗体数量的提高提供了有利条件。实验表明此免疫传感器的线性范围为0.02-80 ng/mL,检出限为0.01ng/mL。3.基于PANI及PDA构建的甲胎蛋白免疫传感器:构建了基于聚苯胺/聚多巴胺(PANI/PDA)检测甲胎蛋白抗原(AFP)的非标记电化学免疫传感器。采用电聚合方法将聚苯胺(PANI)薄膜修饰在玻碳电极表面,然后利用多巴胺(DA)在弱碱性溶液中自聚合为PDA,在PANI表面包覆成膜修饰在电极表面以固定anti-AFP。对聚苯胺的掺杂改性拓宽了其pH适应范围,提高了其在传感器中的应用,此外,聚多巴胺薄膜表面富集了大量的氨基和羟基为该传感器修饰蛋白提供里良好的生物环境。此免疫传感器的线性范围为0.01-80 ng/mL,检出限为0.007 ng/mL。4.基于氮掺杂多孔碳及PoPD构建的甲胎蛋白免疫传感器:利用氮掺杂多孔碳的良好的生物相容性提高了邻苯二胺的电化学活性,增加了AFP的固载数量且保持蛋白的活性,加快电子传递,从而增加免疫传感器的灵敏度。且聚邻苯二胺表面含有很多氨基及亚氨基等,可以提供很多再生基团,为其固载anti-AFP提供了优良条件。在优化条件下,免疫传感器的检测范围为0.005-100ng/mL,检出限为0.0006 ng/mL。
[Abstract]:The research shows that the only important sign can be used to screening and detection of tumor markers is in human serum alpha fetoprotein (AFP) content, timely detection and diagnosis of tumor can accurately and sensitively detect hemorrhage AFP content in Qing Dynasty. The traditional immune monitoring method by analysis of long, expensive and needs analysis instruments the operation of professional personnel and limited its development. This has the advantages of simple operation, high sensitivity, immune sensor for determination of shorter and low cost has been a strong concern of the researchers. According to the different electrode modification material for electrochemical immunosensor with high sensitivity and stable performance were prepared. The electrochemical immunosensor in our main study on the modification is sensitive interface electrically active material. We used in this paper is not easy to be lost in the electrode surface, easy to be fixed on the electrode surface guide The electric conductive polymer. The polymer is widely used in the sensor is mainly due to its excellent properties (environmental friendliness, high conductivity, good stability etc.). Conductive polymers such as polyaniline (PANI), polypyrrole (PPy), poly benzene amine two (PoPD) after doping conductive performance has greatly improved the performance of electrochemical immunosensor, this type of material can be greatly improved. In recent years, biomimetic nano materials because of its excellent film-forming properties, simple preparation and characteristics: good biocompatibility, become the focus of research in the development of immune sensor. Two order reaction platform polydopamine at present, as a kind of good has aroused extensive attention, this is mainly due to the structure of poly dopamine containing amino and phenolic hydroxyl groups, making it a good biocompatibility, is the current number of bio friendly A type of large molecules. This paper uses electro polymerization and method of electro deposition of conductive polymer immobilized on the electrode in order to construct a series of highly sensitive stability and excellent electrochemical immunosensor; test by electrochemical analysis technique on immune electrode self-assembly process and electrode interface were characterized and discussed immunoelectrode application in biological analysis. The main contents of this dissertation are as follows: 1. AFP immunosensor based on PANI doped: assembled with high selectivity, stability, novel label free amperometric immunosensor sensitivity, mainly used for alpha fetoprotein (AFP) detection. Polymerization of polyaniline films modified on the surface of glassy carbon electrode using electricity, then the alpha fetoprotein antibody (anti-AFP) in surface modification of polyaniline. The experiment shows that the immune sensor linear range 0.01-50 ng/mL, the detection limit can reach 0.008 ng/mL. the immune sensor in the preparation process showed high selectivity, reproducibility and stability of the advantages of.2. based on the AFP immunosensor of PPy and PDA Construction: polypyrrole / polydopamine (PPy/PDA) label free electrochemical immunosensor successfully assembled, can be used for the detection of alpha fetoprotein (AFP). By using the method of electro deposition of pyrrole (Py) deposited on the surface of glassy carbon electrode polypyrrole (PPy) film, and then use the dopamine (DA) can self oxidation polymerization in alkaline solution, the PPy coating film on the electrode surface modified with fixed anti-AFP. polydopamine film excellent biocompatibility provides favorable conditions for the number of sensor fixing antibodies increased. Experiments show that the linear range of the immunosensor for 0.02-80 ng/mL, the detection limit is 0.01ng/mL.3. based on PANI and PDA construction AFP immunosensor: Construction of the polyaniline / poly dopamine (PANI/PDA) based on the detection of alpha fetoprotein (AFP) antigen label free electrochemical immunosensor. Polymerization method using electric polyaniline (PANI) film modified on glassy carbon electrode surface, and then use the dopamine (DA) in alkaline solution to self polymerization of PDA. On the surface of PANI film coated on the electrode surface modified with anti-AFP. fixation of polyaniline doped pH to broaden its scope, improve its application in sensors, in addition, the polydopamine film surface enrichment of a large number of amino and hydroxyl groups for the sensor protein modification of good environment. The linear range of the organisms immune the sensor is 0.01-80 ng/mL, a detection limit of 0.007 ng/mL.4. AFP immunosensor based on nitrogen doped porous carbon and PoPD Construction: the use of nitrogen doped porous carbon with good biocompatibility To improve the electrochemical activity of benzene amine two, increase the number of supported and maintained the activity of the protein AFP, accelerate the electron transfer, thereby increasing the sensitivity of the immunosensor. And the poly phthalate two amine surface contains many amino and imino groups, can provide a lot of regeneration, provides excellent conditions for the immobilization of anti-AFP. Under the optimized conditions, the detection range of immune sensor is 0.005-100ng/mL, the detection limit was 0.0006 ng/mL.

【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O657.1

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