本文選題：表面增強拉曼光譜(SERS) + 免疫分析方法(IA)　； 參考：《蘇州大學》2016年碩士論文

【摘要】：表面增強拉曼光譜(Surface-Enhanced Raman Scattering,SERS)因其高靈敏度等優(yōu)勢被廣泛應(yīng)用于化學和生物的檢測分析中,而基于抗原抗體特異性反應(yīng)的免疫分析方法(Immunoassay,IA),其特異性強。SERS結(jié)合IA作為一種新型的檢測手段,兼具了SERS的高靈敏度和免疫分析方法的高特異性,在最近幾年被廣泛應(yīng)用于食品安全檢測及環(huán)境監(jiān)測等領(lǐng)域。在SERS-IA中,用于支撐免疫反應(yīng)的基底研究一直以來都是人們研究的熱點,基底材料各有千秋。本研究的一個主要目的旨在探索出一種能很好支撐免疫反應(yīng)、信號均勻、操作簡便、成本較低的固相基底材料。在幾經(jīng)嘗試之后,發(fā)現(xiàn)硝酸纖維素膜(即NC膜)兼具這些優(yōu)點。在此基礎(chǔ)上,建立了基于NC膜的多通道SERS-IA,并應(yīng)用于肉樣中一種新型β-激動劑(即瘦肉精)溴布特羅(Brombuterol)的檢測。另外,本論文也對以GNPs為免疫探針基底的表面增強拉曼光譜結(jié)合免疫層析技術(shù)(SERS-ICT)進行了深入研究,建立了測定Hg~(2+)的SERS-ICT,并對加標水樣(河水、湖水、自來水)、尿樣、血樣中的Hg~(2+)進行了檢測,取得了令人滿意的結(jié)果。本研究主要內(nèi)容為以下三個部分:一.介紹論文的研究背景及意義。闡述了表面增強拉曼光譜的相關(guān)概念以及應(yīng)用,說明了免疫分析的基本理論,著重介紹了SERS的基底材料,SERS-IA以及SERS-ICT。另外,也敘述了一直以來人們關(guān)注的環(huán)境和食品安全問題。二.在探索以銀膜、銀納米顆粒、金納米顆粒作為免疫反應(yīng)的支撐物時,發(fā)現(xiàn)這些材料存在前處理和制備過程復雜繁瑣,與蛋白結(jié)合不牢固,而且重現(xiàn)性差,信號不均勻等種種問題。而以NC膜為免疫反應(yīng)的固相支撐物,不僅與蛋白的交聯(lián)作用好,而且快速、方便。與此同時制備了一種基于新型核殼型GNPs@Ag納米粒子結(jié)合抗體的具有拉曼活性的免疫探針,建立了基于SERS和NC膜的免疫分析方法(SERS-IA),并對溴布特羅進行檢測,IC50和檢出限分別為0.02 ng m L~(-1)和0.16 pg m L~(-1),顯示出了高的靈敏性。由于溴布特羅與鹽酸克倫特羅的分子結(jié)構(gòu)相近,SERS-IA與鹽酸克倫特羅存在40.8%的交叉反應(yīng)。SERS-IA與其他β-激動劑無交叉反應(yīng)。將建立的SERS-IA應(yīng)用于加標肉樣中溴布特羅的檢測,結(jié)果令人滿意。三.本論文對表面增強拉曼光譜結(jié)合免疫層析技術(shù)(SERS-ICT)進行了深入研究。制備了以Au NPs為基底的SERS免疫探針,建立了測定Hg~(2+)的SERS-ICT,對加標水樣(河水、湖水、自來水)、尿樣、血樣中的Hg~(2+)進行了檢測。在最優(yōu)的實驗條件下,該SERS-ICT有著低于ng g~(-1)級的高靈敏度,IC50和檢出限LOD分別為0.12 ng m L~(-1)和0.45 pg m L~(-1)。與19種陰、陽離子的交叉反應(yīng)率均小于0.01%。另外,對與汞接觸的職業(yè)病患者實際樣品中Hg~(2+)進行了檢測,并與其他方法進行了比較,取得了令人滿意的結(jié)果。
[Abstract]:Surface-enhanced Raman scattering spectroscopy (SERS) is widely used in chemical and biological analysis because of its high sensitivity. As a new detection method, Immunoassay based on specific reaction of antigen and antibody (Immunoassay IA) has both the high sensitivity of SERS and the high specificity of immunoassay. In recent years, it has been widely used in food safety detection and environmental monitoring. In SERS-IA, substrate research for supporting immune response has been a hot topic, and substrate materials have their own merits. One of the main purposes of this study is to explore a solid phase substrate material which can support immune response well, signal is uniform, operation is simple and cost is low. After several attempts, it was found that the nitrocellulose membrane (NC membrane) had these advantages. On this basis, a multichannel SERS-IA-based NC membrane was established and applied to the detection of brombuterol, a new 尾 -agonist (lean meatine) in meat samples. In addition, the surface enhanced Raman spectroscopy combined with immunochromatography (SERS-ICT) based on GNPs was also studied in this paper. The SERS-ICTs for the determination of Hg2 were established, and the standard water samples (river, lake, tap water), urine samples were also studied. The detection of Hg ~ (2) in blood samples has been carried out and satisfactory results have been obtained. The main contents of this study are as follows: 1. This paper introduces the research background and significance. The related concepts and applications of surface-enhanced Raman spectroscopy are described. The basic theory of immunoassay is explained. The substrate materials of SERS, SERS-IA and SERS-ICTare emphatically introduced. In addition, it also describes the environmental and food safety issues that people have been concerned about all along. II. When exploring silver film, silver nanoparticles and gold nanoparticles as the support of immune reaction, it was found that the pretreatment and preparation of these materials were complicated, the binding to proteins was not firm, and the reproducibility of these materials was poor. A variety of problems such as uneven signals. The solid phase support with NC membrane not only crosslinked with protein, but also was fast and convenient. At the same time, a novel Raman immunosurgery probe based on novel core-shell GNPs @ Ag nanoparticles binding antibody was prepared. An immunoassay (SERS-IA) based on SERS and NC membranes was established. The detection limits of IC50 and detection limits for bromobuterol were 0.02 ng mL ~ (-1) and 0.16 PG mL ~ (-1), respectively, showing a high sensitivity. Because the molecular structure of bromobuterol and clenbuterol hydrochloride is similar, there is 40.8% cross reaction between SERS-IA and clenbuterol hydrochloride. There is no cross reaction between SERS-IA and other 尾 -agonists. The established SERS-IA was applied to the determination of bromobuterol in standard meat samples with satisfactory results. III. Surface enhanced Raman spectroscopy combined with immunochromatography (SERS-ICT) has been studied in this paper. A SERS immunoprobe based on au NPs was prepared and a SERS-ICT-based method was established for the determination of Hg2 in water (river, lake, tap water), urine and blood samples. Under the optimal experimental conditions, the SERS-ICT has a high sensitivity IC50 and a detection limit of 0.12 ng mL ~ (-1) and 0.45 PG mL ~ (-1), respectively, which are lower than ng g ~ (-1). The cross reaction rate with 19 kinds of anions and cations was less than 0.01. In addition, Hg ~ (2) in the actual samples of occupational disease patients exposed to mercury were detected and compared with other methods, and satisfactory results were obtained.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：O657.37

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