基于新型復(fù)合納米材料的電致化學(xué)發(fā)光及生物傳感研究
發(fā)布時(shí)間：2018-09-17 16:12
【摘要】：當(dāng)今,隨著納米技術(shù)的快速發(fā)展,納米材料已被廣泛應(yīng)用于ECL生物傳感提高信號響應(yīng)。電致化學(xué)發(fā)光(ECL)和電化學(xué)分析是通過電化學(xué)反應(yīng)引發(fā)的、簡便、靈敏、強(qiáng)有力的分析檢測技術(shù)。電致化學(xué)發(fā)光因其背景信號低、線性范圍寬、靈敏度高、選擇性好、操作簡單以及成本低等優(yōu)點(diǎn),受到越來越多科研工作者的廣泛研究。本文研究制備了多種新型復(fù)合納米材料,并基于目標(biāo)循環(huán)放大原理構(gòu)建了電致化學(xué)發(fā)光和電化學(xué)生物傳感器,實(shí)現(xiàn)了對凝血酶、CEA以及Hg~(2+)的高靈敏檢測。本文主要進(jìn)行了以下幾個(gè)方面的研究:1.通過原位還原制備了一種新型的納米材料AgNCs,該納米材料具有較強(qiáng)的ECL和電化學(xué)性能,通過DNA酶輔助目標(biāo)循環(huán)和HCR雙重放大技術(shù),實(shí)現(xiàn)了凝血酶的高靈敏檢測。目標(biāo)凝血酶存在下打開發(fā)夾DNA,再利用DNA酶催化循環(huán)剪切過量的底物,產(chǎn)生大量基底片段s1。納米金修飾的電極通過SH-DNA連接s1,進(jìn)一步引發(fā)HCR。利用AgNO_3和硼氫化鈉在dsDNA模板修飾的電極表面原位合成大量的AgNCs。通過DNA酶循環(huán)和HCR雙重放大技術(shù),組裝大量的AgNCs,極大地增強(qiáng)了ECL和電化學(xué)信號響應(yīng),實(shí)現(xiàn)了凝血酶的高靈敏檢測。該方法有望應(yīng)用于多種目標(biāo)生物分子的高靈敏檢測。2.首先制備了Ru(bpy)_3~(2+)摻雜SiO_2的新型納米復(fù)合材料(Ru@SiO_2)作為ECL發(fā)光劑,利用三丙胺(TPA)作為ECL共反應(yīng)劑,基于Fc對Ru@SiO_2 ECL信號的有效淬滅,通過目標(biāo)循環(huán)放大技術(shù)研制了新型的電化學(xué)發(fā)光生物傳感器,實(shí)現(xiàn)了對CEA的高靈敏檢測。首先利用核殼結(jié)構(gòu)的磁珠(Au@MBs)固定Ru@Si O2信號探針,再通過DNA雜交技術(shù)連接Fc-DNA,淬滅ECL信號。通過目標(biāo)循環(huán)放大技術(shù)產(chǎn)生大量的DNA1,DNA1取代磁珠上的Fc-DNA,探針的ECL信號增強(qiáng)。該ECL生物傳感器具有良好的穩(wěn)定性和較高的選擇性,檢測范圍為10 fg/m L~10 ng/m L,最低檢測限為2.3 fg/mL。3.首先設(shè)計(jì)了一種非線性雜交鏈?zhǔn)椒磻?yīng),通過引物引發(fā)雙鏈基底DNA自組裝成帶有大量ECL探針的樹枝狀納米結(jié)構(gòu)。利用Exo-III和Nt.BbvCI核酸內(nèi)切酶輔助的循環(huán)放大技術(shù)并結(jié)合樹枝狀納米結(jié)構(gòu),構(gòu)建了ECL生物傳感器用于目標(biāo)Hg~(2+)的高靈敏檢測。該生物傳感器具有較高的選擇性和較低的檢測限,檢測的線性范圍為100 fM~50 nM,檢測限為23.6 fM。該ECL生物傳感器不僅能檢測水樣中的Hg~(2+),在其它目標(biāo)的檢測中也具有良好的應(yīng)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.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.1
，
本文編號：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.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.1
，
本文編號：2246449
本文鏈接：http://sikaile.net/kejilunwen/huaxue/2246449.html
最近更新
教材專著
