基于新型复合纳米材料的电致化学发光及生物传感研究
发布时间：2018-09-17 16:12
【摘要】：当今,随着纳米技术的快速发展,纳米材料已被广泛应用于ECL生物传感提高信号响应。电致化学发光(ECL)和电化学分析是通过电化学反应引发的、简便、灵敏、强有力的分析检测技术。电致化学发光因其背景信号低、线性范围宽、灵敏度高、选择性好、操作简单以及成本低等优点,受到越来越多科研工作者的广泛研究。本文研究制备了多种新型复合纳米材料,并基于目标循环放大原理构建了电致化学发光和电化学生物传感器,实现了对凝血酶、CEA以及Hg~(2+)的高灵敏检测。本文主要进行了以下几个方面的研究:1.通过原位还原制备了一种新型的纳米材料AgNCs,该纳米材料具有较强的ECL和电化学性能,通过DNA酶辅助目标循环和HCR双重放大技术,实现了凝血酶的高灵敏检测。目标凝血酶存在下打开发夹DNA,再利用DNA酶催化循环剪切过量的底物,产生大量基底片段s1。纳米金修饰的电极通过SH-DNA连接s1,进一步引发HCR。利用AgNO_3和硼氢化钠在dsDNA模板修饰的电极表面原位合成大量的AgNCs。通过DNA酶循环和HCR双重放大技术,组装大量的AgNCs,极大地增强了ECL和电化学信号响应,实现了凝血酶的高灵敏检测。该方法有望应用于多种目标生物分子的高灵敏检测。2.首先制备了Ru(bpy)_3~(2+)掺杂SiO_2的新型纳米复合材料(Ru@SiO_2)作为ECL发光剂,利用三丙胺(TPA)作为ECL共反应剂,基于Fc对Ru@SiO_2 ECL信号的有效淬灭,通过目标循环放大技术研制了新型的电化学发光生物传感器,实现了对CEA的高灵敏检测。首先利用核壳结构的磁珠(Au@MBs)固定Ru@Si O2信号探针,再通过DNA杂交技术连接Fc-DNA,淬灭ECL信号。通过目标循环放大技术产生大量的DNA1,DNA1取代磁珠上的Fc-DNA,探针的ECL信号增强。该ECL生物传感器具有良好的稳定性和较高的选择性,检测范围为10 fg/m L~10 ng/m L,最低检测限为2.3 fg/mL。3.首先设计了一种非线性杂交链式反应,通过引物引发双链基底DNA自组装成带有大量ECL探针的树枝状纳米结构。利用Exo-III和Nt.BbvCI核酸内切酶辅助的循环放大技术并结合树枝状纳米结构,构建了ECL生物传感器用于目标Hg~(2+)的高灵敏检测。该生物传感器具有较高的选择性和较低的检测限,检测的线性范围为100 fM~50 nM,检测限为23.6 fM。该ECL生物传感器不仅能检测水样中的Hg~(2+),在其它目标的检测中也具有良好的应用前景。
[Abstract]:Nowadays, with the rapid development of nanotechnology, nanomaterials have been widely used in ECL biosensor to improve signal response. Electrochemiluminescence (ECL) and electrochemical analysis are simple, sensitive and powerful analytical techniques initiated by electrochemical reaction. Electrochemiluminescence is widely studied by more and more researchers because of its low background signal, wide linear range, high sensitivity, good selectivity, simple operation and low cost. In this paper, a variety of novel composite nanomaterials were prepared, and electrochemiluminescence and electrochemical biosensors were constructed based on the principle of target cyclic amplification. The highly sensitive detection of Hg~ _ (2) and Hg~ _ (2) were achieved. This article mainly carries on the following several aspects of research: 1. A new nano-material, AgNCs, was prepared by in-situ reduction. The nano-material has strong ECL and electrochemical properties. The high sensitivity detection of thrombin was realized by means of DNA enzyme assisted target cycle and HCR double amplification technique. In the presence of target thrombin, DNA, was opened and DNA was used to catalyze the cyclic shearing of excessive substrates, resulting in a large number of substrates S1. Nanocrystalline gold modified electrode is connected to S1 by SH-DNA, which further initiates HCR.. In situ Synthesis of large amounts of AgNCs. on the Surface of the electrode modified by dsDNA template using AgNO_3 and Sodium Borohydride By means of DNA enzyme cycle and HCR double amplification technique, a large number of AgNCs, were assembled to greatly enhance the response of ECL and electrochemical signal, and the high sensitivity detection of thrombin was realized. This method is expected to be applied to the detection of many target biomolecules. Firstly, a new type of SiO_2 doped nanocomposites (Ru@SiO_2) with Ru (bpy) _ 3 ~ (2) -doped SiO_2 was prepared as ECL luminescent agent. Tripropylamine (TPA) was used as ECL co-reaction agent to quench Ru@SiO_2 ECL signal effectively based on Fc. A novel electrochemiluminescence biosensor was developed by means of target cyclic amplification, which can detect CEA with high sensitivity. Firstly, the Ru@Si O 2 signal probe was fixed by magnetic beads (Au@MBs) with core-shell structure, and then the Fc-DNA, quenched ECL signal was connected by DNA hybridization technique. A large number of ECL signals are enhanced by DNA1,DNA1 instead of Fc-DNA, probe on magnetic beads by target cyclic amplification technique. The ECL biosensor has good stability and high selectivity. The detection range is 10 fg/m L ~ (10) ng/m / L and the minimum detection limit is 2.3 fg/mL.3.. A nonlinear hybrid chain reaction was designed to initiate the self-assembly of double-stranded substrates DNA into dendritic nanostructures with a large number of ECL probes. Using Exo-III and Nt.BbvCI endonuclease assisted cyclic amplification techniques and dendritic nanostructures, a ECL biosensor was constructed to detect target Hg~ (2) with high sensitivity. The biosensor has high selectivity and low detection limit. The linear range of the biosensor is 100 fM~50 nM, and the detection limit is 23.6 fM.. The ECL biosensor not only can detect Hg~ (2) in water samples, but also has a good application prospect in other target detection.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1
，
本文编号：2246449
[Abstract]:Nowadays, with the rapid development of nanotechnology, nanomaterials have been widely used in ECL biosensor to improve signal response. Electrochemiluminescence (ECL) and electrochemical analysis are simple, sensitive and powerful analytical techniques initiated by electrochemical reaction. Electrochemiluminescence is widely studied by more and more researchers because of its low background signal, wide linear range, high sensitivity, good selectivity, simple operation and low cost. In this paper, a variety of novel composite nanomaterials were prepared, and electrochemiluminescence and electrochemical biosensors were constructed based on the principle of target cyclic amplification. The highly sensitive detection of Hg~ _ (2) and Hg~ _ (2) were achieved. This article mainly carries on the following several aspects of research: 1. A new nano-material, AgNCs, was prepared by in-situ reduction. The nano-material has strong ECL and electrochemical properties. The high sensitivity detection of thrombin was realized by means of DNA enzyme assisted target cycle and HCR double amplification technique. In the presence of target thrombin, DNA, was opened and DNA was used to catalyze the cyclic shearing of excessive substrates, resulting in a large number of substrates S1. Nanocrystalline gold modified electrode is connected to S1 by SH-DNA, which further initiates HCR.. In situ Synthesis of large amounts of AgNCs. on the Surface of the electrode modified by dsDNA template using AgNO_3 and Sodium Borohydride By means of DNA enzyme cycle and HCR double amplification technique, a large number of AgNCs, were assembled to greatly enhance the response of ECL and electrochemical signal, and the high sensitivity detection of thrombin was realized. This method is expected to be applied to the detection of many target biomolecules. Firstly, a new type of SiO_2 doped nanocomposites (Ru@SiO_2) with Ru (bpy) _ 3 ~ (2) -doped SiO_2 was prepared as ECL luminescent agent. Tripropylamine (TPA) was used as ECL co-reaction agent to quench Ru@SiO_2 ECL signal effectively based on Fc. A novel electrochemiluminescence biosensor was developed by means of target cyclic amplification, which can detect CEA with high sensitivity. Firstly, the Ru@Si O 2 signal probe was fixed by magnetic beads (Au@MBs) with core-shell structure, and then the Fc-DNA, quenched ECL signal was connected by DNA hybridization technique. A large number of ECL signals are enhanced by DNA1,DNA1 instead of Fc-DNA, probe on magnetic beads by target cyclic amplification technique. The ECL biosensor has good stability and high selectivity. The detection range is 10 fg/m L ~ (10) ng/m / L and the minimum detection limit is 2.3 fg/mL.3.. A nonlinear hybrid chain reaction was designed to initiate the self-assembly of double-stranded substrates DNA into dendritic nanostructures with a large number of ECL probes. Using Exo-III and Nt.BbvCI endonuclease assisted cyclic amplification techniques and dendritic nanostructures, a ECL biosensor was constructed to detect target Hg~ (2) with high sensitivity. The biosensor has high selectivity and low detection limit. The linear range of the biosensor is 100 fM~50 nM, and the detection limit is 23.6 fM.. The ECL biosensor not only can detect Hg~ (2) in water samples, but also has a good application prospect in other target detection.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1
，
本文编号：2246449
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