金仿生矿化复合酶的制备及其初步的医学与环境应用
发布时间：2018-11-15 21:02
【摘要】：贵金属纳米簇尤其金纳米簇(Au NCs)以其优良的尺寸效应和光电功能优势,被广泛应用于生物标记、酶催化、成像分析、以及传感检测等领域。本文采用生物仿生矿化途径,一方面制备了一种金矿化的葡萄糖氧化酶(GOx)复合物,用以修饰核酸探针并结合磁分离技术,发展了一种电化学信号放大的夹心法,用于炭疽病毒DNA序列的超灵敏检测及其单碱基突变分析;另一方面,开发了一种血红素(Hemin)介导金矿化的多功能模拟酶,用以标记甲胎蛋白(AFP)抗体,并借助其具有的过氧化物酶催化与金催化银沉积功能,实现了对人血清中AFP的快速、灵敏、可视化检测;与此同时,将该模拟酶与Fe3O4负载的三聚氰胺甲醛树脂(m PMF)介孔聚合物相结合,开发了一种可回收利用的多功能复合材料,用于环境有机毒物(染料)的高效吸附与催化降解。主要内容包括:(1)建立一种基于金仿生矿化GOx的电化学传感技术,应用于炭疽病毒DNA序列的检测及其单碱基突变分析(第二章)。一方面,以GOx蛋白为还原剂和稳定剂,通过原位金仿生矿化途径制得GOx-Au复合酶,用以标记经酪胺修饰的核酸检测探针,同时,以磁性颗粒负载经酪胺修饰的核酸捕获探针,进而将二者与目标链杂交形成“三明治”型核酸复合物;另一方面,借助GOx-Au复合酶中GOx催化葡萄糖分解得到双氧水,进而利用其中矿化金具有的类过氧化物酶功能催化双氧水氧化酪胺偶联,籍以连接DNA杂交反应后的捕获探针和检测探针,并借助金催化金沉积途径放大其电化学信号,建立了一种电化学传感新技术,实现了对血液中炭疽病毒DNA的超灵敏检测及其单碱基突变分析。由此建立的基于金仿生矿化GOx催化信号放大途径的电化学传感方法,突破了低含量短链DNA的检测瓶颈,可望为临床炭疽病毒感染的早期诊断提供一种新的检测工具。(2)基于血红素介导金矿化途径的模拟酶复合物的制备及其用于AFP的比色分析(第三章)。采用生物矿化途径,以Hemin为模板于碱性条件下介导金矿化,制得Hemin-Au模拟酶复合物。研究表明,Hemin-Au兼具Hemin类过氧化物酶和金的双重催化功能。以肝癌标志物AFP的免疫分析为例,将该模拟酶复合物标记AFP抗体,并结合酶联免疫吸附试验途径,建立了酶催化底物显色和金催化银沉积反应两种信号放大途径,用以分别实现了对血液中AFP的高灵敏比色检测。(3)基于磁性颗粒负载m PMF介孔聚合物的模拟酶复合材料的制备及其用于环境染料的吸附与催化降解(第四章)。采用水热法合成磁性纳米颗粒,进而借助三聚氰胺和甲醛的羟基化缩聚反应形成m PMF介孔聚合物,并包覆于磁性纳米颗粒表面,制得Fe3O4@m PMF磁性复合物。然后,通过化学键合方法将Hemin-Au模拟酶负载于Fe3O4@m PMF颗粒上,制得具有高比表面积、强催化功能、可回收利用的多功能复合材料。以亚甲基蓝(MB)和罗丹明B(RhB)为例,系统地考察了该复合材料吸附与催化降解常见环境染料的性能。结果表明,这种基于mPMF介孔聚合物的磁性模拟酶复合材料,可望作为一种绿色、高效的吸附与催化新材料,广泛应用于染料等环境毒物的高效处理。
[Abstract]:The noble metal nanoclusters, especially the gold nanoclusters (Au NCs), are widely used in the fields of biological markers, enzyme catalysis, imaging analysis and sensing detection. In this paper, a gold-gold-based glucose oxidase (GOx) complex is prepared by means of biomimetic mineralization, which is used to modify the nucleic acid probe and combine the magnetic separation technology to develop a sandwich method for electrochemical signal amplification. The invention relates to an ultra-sensitive detection of a DNA sequence of a anthrax virus and a single-base mutation analysis thereof, and on the other hand, a multi-functional analog enzyme for mediating a gold mineralization of a hemin is developed to label the alpha-fetoprotein (AFP) antibody, The method realizes the rapid, sensitive and visual detection of the AFP in human serum by virtue of the peroxidase catalysis and the gold catalytic silver deposition function, The multifunctional composite material can be recycled, and is used for high-efficiency adsorption and catalytic degradation of the environment organic poison (dye). The main contents include: (1) To establish an electrochemical sensing technology based on the gold biomimetic mineralization GOx, which is applied to the detection of the DNA sequence of the anthrax virus and the single base mutation analysis (the second chapter). in one aspect, a GOx-Au composite enzyme is prepared by a GOx protein as a reducing agent and a stabilizing agent through an in-situ gold biomimetic mineralization route, then, the two are hybridized with the target chain to form a 鈥渟andwich鈥，
本文编号：2334451
[Abstract]:The noble metal nanoclusters, especially the gold nanoclusters (Au NCs), are widely used in the fields of biological markers, enzyme catalysis, imaging analysis and sensing detection. In this paper, a gold-gold-based glucose oxidase (GOx) complex is prepared by means of biomimetic mineralization, which is used to modify the nucleic acid probe and combine the magnetic separation technology to develop a sandwich method for electrochemical signal amplification. The invention relates to an ultra-sensitive detection of a DNA sequence of a anthrax virus and a single-base mutation analysis thereof, and on the other hand, a multi-functional analog enzyme for mediating a gold mineralization of a hemin is developed to label the alpha-fetoprotein (AFP) antibody, The method realizes the rapid, sensitive and visual detection of the AFP in human serum by virtue of the peroxidase catalysis and the gold catalytic silver deposition function, The multifunctional composite material can be recycled, and is used for high-efficiency adsorption and catalytic degradation of the environment organic poison (dye). The main contents include: (1) To establish an electrochemical sensing technology based on the gold biomimetic mineralization GOx, which is applied to the detection of the DNA sequence of the anthrax virus and the single base mutation analysis (the second chapter). in one aspect, a GOx-Au composite enzyme is prepared by a GOx protein as a reducing agent and a stabilizing agent through an in-situ gold biomimetic mineralization route, then, the two are hybridized with the target chain to form a 鈥渟andwich鈥，
本文编号：2334451
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