本文關(guān)鍵詞： 谷胱甘肽 電化學(xué)阻抗 DNA 谷胱甘肽還原酶 檢測(cè)　出處：《中南林業(yè)科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：電化學(xué)傳感器由于其選擇性好,檢測(cè)耗時(shí)短,無污染等優(yōu)點(diǎn),在發(fā)酵工業(yè),疾病預(yù)防,醫(yī)藥食品加工等相關(guān)領(lǐng)域具有廣泛的應(yīng)用前景。而納米材料由于具有良好的比表面積和獨(dú)特的導(dǎo)電性能,被應(yīng)用到高度敏感的生物感應(yīng)方法設(shè)計(jì)中,如石墨烯,量子點(diǎn),金納米粒子及介孔二氧化硅納米顆粒等。本論文針對(duì)谷胱甘肽(GSH)研究了一系列成本低、快速檢測(cè)、具有更寬檢測(cè)范圍的新型電化學(xué)生物傳感器,其主要內(nèi)容如下:(1)DNA門控開關(guān)功能化介孔二氧化硅納米粒子電化學(xué)檢測(cè)GSH采用介孔二氧化硅粒子和DNA門控系統(tǒng)設(shè)計(jì)一種基于正十二硫醇修飾絲網(wǎng)印刷電極(SPE)的直接快速檢測(cè)GSH的電化學(xué)方法。用于電化學(xué)檢測(cè)的T-Hg2+-T結(jié)構(gòu)的DNA門控系統(tǒng)可特異性與GSH發(fā)生取代反應(yīng),門控系統(tǒng)被打開,釋放出亞甲基藍(lán)分子。然后,由于亞甲基藍(lán)(MB)分子滲透到硫醇層堆積,被固定的十二硫醇不僅能避免蛋白的非特異性吸附也提高電流響應(yīng)信號(hào),其檢測(cè)范圍1.0 nmol·L-1～1.0mmol·L-1,檢測(cè)下限為0.6nmol·L-1。該方法檢測(cè)下限較低,不用對(duì)檢測(cè)樣品處理,操作簡(jiǎn)單。(2)Hg2+觸發(fā)鏈?zhǔn)诫s交反應(yīng)構(gòu)建電化學(xué)傳感器檢測(cè)谷胱甘肽是基于雜交鏈?zhǔn)椒磻?yīng)信號(hào)放大,我們?cè)O(shè)計(jì)一個(gè)具有高度敏感性、簡(jiǎn)單的GSH電化學(xué)檢測(cè)方法。通過對(duì)目標(biāo)分子進(jìn)行雜交鏈?zhǔn)椒磻?yīng)在傳感器表面形成dsDNA聚合物。指示劑可以有效地嵌入到雙鏈DNA聚合物凹槽處,從而實(shí)現(xiàn)對(duì)GSH的檢測(cè),該方法的對(duì)數(shù)線性響應(yīng)范圍為1～500nmol·L-1,檢測(cè)下限可達(dá)0.6nmol·L-1。該法技術(shù)已被證明是一種簡(jiǎn)單且高度敏感的對(duì)不同生物分子具有檢測(cè)作用的信號(hào)轉(zhuǎn)導(dǎo)途徑。(3)電化學(xué)阻抗法快速檢測(cè)GSH及谷胱甘肽還原酶(GR)是金電極經(jīng)過GSH修飾后,其阻抗值顯著增加,而阻抗的增加與GSH濃度密切相關(guān)。通過檢測(cè)酶反應(yīng)溶液中GSH的變化,我們可以間接地求得GR的活性。該傳感器對(duì)GSH的檢測(cè)范圍為250μmol·L-1-16 mmol·L-1。檢測(cè)酶活性范圍在0.005-0.5 U。此外,通過與比色法檢測(cè)得到的GR活性進(jìn)行比較,結(jié)果表明電化學(xué)檢測(cè)方法具有良好的可靠性。與其他檢測(cè)方法相比,這種方法具有靈敏度高、耗時(shí)較少的優(yōu)點(diǎn)。
[Abstract]:Because of its good selectivity, short detection time, no pollution and other advantages, electrochemical sensors in the fermentation industry, disease prevention. Because of its good specific surface area and unique electrical conductivity, nano-materials have been used in the design of highly sensitive biological sensing methods, such as graphene. Quantum dots, gold nanoparticles and mesoporous silica nanoparticles. In this paper, a series of low-cost and rapid detection of glutathione glutathione (GSH) have been studied. A novel electrochemical biosensor with a wider detection range. The main contents are as follows:. Electrochemical detection of DNA gated switch functionalized mesoporous silica nanoparticles GSH based on 12 mercaptan modified screen printing electrode was designed using mesoporous silica particles and DNA gating system. The T-Hg2-T DNA gating system for electrochemical detection can react specifically with GSH. The gated system is opened to release methylene blue molecules, which then penetrate into the mercaptan layer due to methylene blue MBs. The immobilized 12 mercaptan could not only avoid the non-specific adsorption of the protein, but also enhance the current response signal. The detection range was 1.0 nmol 路L ~ (-1) ~ (-1) ~ (-1) nmol 路L ~ (-1). The detection limit is 0.6 nmol 路L ~ (-1). Simple operation. Hg2 triggered chain hybridization construction of electrochemical sensor for detection of glutathione is based on the amplification of hybrid chain reaction signal, we design a highly sensitive one. Simple GSH electrochemical detection method. DsDNA polymer is formed on the sensor surface by hybrid chain reaction of the target molecule. The indicator can be effectively embedded in the groove of double-stranded DNA polymer. The logarithmic linear response range of the method is 1 ~ 500nmol 路L ~ (-1). The detection limit can reach 0.6 nmol 路L ~ (-1). This method has been proved to be a simple and highly sensitive signal transduction pathway for different biomolecules. Electrochemical impedance method for rapid detection of GSH and glutathione reductase (Glutathione reductase) is a gold electrode modified by GSH. The impedance value was significantly increased, and the increase of impedance was closely related to the concentration of GSH. The changes of GSH in enzymatic reaction solution were detected. The activity of gr can be obtained indirectly. The detection range of GSH by this sensor is 250 渭 mol 路L-1-16. Mmol 路L ~ (-1). The range of enzyme activity was 0.005-0.5 U. in addition. The comparison of gr activity with colorimetric method shows that the electrochemical detection method has good reliability, and compared with other detection methods, this method has the advantages of high sensitivity and less time consuming.
【學(xué)位授予單位】：中南林業(yè)科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TP212;Q503
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本文編號(hào)：1451572