本文選題：電化學(xué)發(fā)光 + 電化學(xué)發(fā)光免疫傳感器�。� 參考：《西南大學(xué)》2015年碩士論文

【摘要】：電化學(xué)發(fā)光(ECL)分析技術(shù)集合了電化學(xué)可控性強(qiáng)和發(fā)光分析靈敏度高的優(yōu)點(diǎn),是一種高的痕量分析檢測方法。電化學(xué)發(fā)光免疫分析法(ECLIA)是電化學(xué)發(fā)光分析和免疫相結(jié)合的產(chǎn)物。這種方法基于抗原抗體之間的特異性免疫反應(yīng)識別目標(biāo)物,建立發(fā)光物質(zhì)與目標(biāo)物之間的定量聯(lián)系,通過測定發(fā)光物質(zhì)的ECL信號的改變,實(shí)現(xiàn)目標(biāo)檢測物的高選擇性、高靈敏度檢測。電化學(xué)發(fā)光免疫傳感器是生物傳感器的一種,其原理是將抗原/抗體固定在電極上,以電化學(xué)發(fā)光信號強(qiáng)度的變化對抗原/抗體進(jìn)行測定。電化學(xué)發(fā)光免疫傳感器具有背景低、靈敏度高、線性范圍寬、時(shí)間/空間可控性好、儀器簡單、分析速度快等優(yōu)點(diǎn),因此,引起了研究者們廣泛興趣�，F(xiàn)已經(jīng)發(fā)展成為生物分析檢測領(lǐng)域重要研究手段,可檢測不同分子量大小的抗體、抗原和半抗原。與熒光能量轉(zhuǎn)移原理類似,電化學(xué)發(fā)光物質(zhì)同樣可以作為能量供體光源而發(fā)生能量轉(zhuǎn)移。在電化學(xué)發(fā)光能量轉(zhuǎn)移中(ECL-ET),通過在電極表面施加一定的電位,能量供體光源在電極上發(fā)生電化學(xué)反應(yīng)產(chǎn)生電生物質(zhì),電生物質(zhì)之間或電生物質(zhì)與溶液中某些組分發(fā)生化學(xué)反應(yīng)而躍遷到激發(fā)態(tài),最后將能量轉(zhuǎn)移給受體而淬滅。釕聯(lián)吡啶(Ru(bpy)32+)及其衍生物具有發(fā)光效率高、可電化學(xué)再生、水溶性好、化學(xué)性能穩(wěn)定等諸多優(yōu)點(diǎn),是電化學(xué)發(fā)光領(lǐng)域應(yīng)用最為廣泛的體系之一。本文將Ru(bpy)32+作為能量供體光源應(yīng)用到ECL-ET體系中,構(gòu)建了新型一次性電化學(xué)發(fā)光免疫傳感器。具體工作如下：利用蠟燭灰通過簡單的方法合成具有高效猝滅效率的無定形碳納米粒子(ACNPs),研究中發(fā)現(xiàn)ACNPs能夠淬滅Ru(bpy)32+電化學(xué)發(fā)光,基于此現(xiàn)象將其用于檢測模型蛋白(MIgG)。先將ACNPs標(biāo)記MIgG,制備ACNP-MIgG納米復(fù)合物,然后與電極表面上Ru(bpy)32+標(biāo)記的抗原發(fā)生免疫反應(yīng)。借助特異性免疫反應(yīng)拉近ACNPs與Ru(bpy)32+之間距離,引發(fā)高效的ECL-ET過程。當(dāng)樣品中存在游離MIgG時(shí),能夠與ACNP-MIgG發(fā)生競爭性免疫反應(yīng),Ru(bpy)32+標(biāo)記的抗體所捕獲的ACNP-MIgG減少,電化學(xué)發(fā)光猝滅率降低,進(jìn)而實(shí)現(xiàn)對MIgG定量分析。所得線性范圍為0.5-400 ng/mL,檢測限為0.35 ng/mL,低于酶聯(lián)免疫方法的3.2 μg/L,穩(wěn)定性、特異性和加標(biāo)回收率均令人滿意。一方面,該方法不需要外在激發(fā)光源,干擾少,背景低；另一方面,與使用電化學(xué)發(fā)光物質(zhì)作為能量受體相比,不會(huì)出現(xiàn)發(fā)射光譜的重疊。該方法操作簡單,成本低廉,具有較高的靈敏度和特異性,適用于床前測試和現(xiàn)場分析,顯示出了較大的應(yīng)用前景。
[Abstract]:Electrochemiluminescence (ECL) analysis technology combines the advantages of high electrochemical controllability and high luminescence analysis sensitivity. It is a high trace analysis method. Electrochemical luminescence immunoassay (ECLIA) is a combination of electrochemiluminescence analysis and immunoassay. This method is based on the specific immune reaction between antigen and antibody to identify the target, establish the quantitative relationship between the luminous substance and the target, and achieve the high selectivity of the target detection by measuring the change of ECL signal of the luminous substance. High sensitivity detection. Electrochemiluminescence immunosensor is a kind of biosensor whose principle is to immobilize antigen / antibody on electrode and measure antigen / antibody with the change of ECL signal intensity. Electrochemiluminescence immunosensor has the advantages of low background, high sensitivity, wide linear range, good time / space controllability, simple instrument and fast analytical speed. It has been developed into an important research tool in the field of biological analysis, which can detect antibodies, antigens and haptens of different molecular weight. Similar to the principle of fluorescence energy transfer, electrochemiluminescence materials can also be used as energy source for energy transfer. In the process of electroluminescent energy transfer, by applying a certain potential on the surface of the electrode, the energy donor light source produces electrochemistry reaction on the electrode to produce electric biomass. The chemical reaction between the electric biomass and some components of the solution leads to the transition to the excited state, and finally the energy is transferred to the receptor and quenched. Ruthenium bipyridyl Rubpyridyl 32) and its derivatives have many advantages, such as high luminescence efficiency, electrochemical regeneration, good water solubility, stable chemical properties and so on. It is one of the most widely used systems in the field of electrochemical luminescence. In this paper, a new type of one-off electrochemical luminescent immunosensor was constructed by using Rubpy32 as a source of energy donor in ECL-ET system. The main work is as follows: the amorphous carbon nanoparticles (ACNPsN) with high quenching efficiency were synthesized by a simple method using candle ash. It was found that ACNPs can quench the electrochemiluminescence (ECL) of Rubpy32. Based on this phenomenon, the ACNPs can be used to detect the model protein. ACNP-MIgG nanocomposites were prepared by labeling ACNPs with MIgG, and then reacted with Rubpy32 labeled antigen on the electrode surface. The distance between ACNPs and RubpyN 32 was narrowed by specific immunoreaction, which induced an efficient ECL-ET process. In the presence of free MIgG in the sample, the ACNP-MIgG could be competitively reacted with ACNP-MIgG. The capture of ACNP-MIgG by the antibody labeled with Rubpyr-32 decreased, and the electrochemiluminescence quenching rate decreased, thus the quantitative analysis of MIgG was realized. The linear range was 0.5-400 ng / mL, and the detection limit was 0.35 ng / mL, which was lower than 3.2 渭 g / L of Elisa. The stability, specificity and recovery were satisfactory. On the one hand, the method does not need external excitation light source, so it has less interference and low background. On the other hand, compared with the use of electroluminescent material as energy receptor, there is no overlap of emission spectra. The method is simple in operation, low in cost, high in sensitivity and specificity. It is suitable for bed test and field analysis, and shows great application prospect.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.6

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