本文選題：熒光 + 金納米簇��； 參考：《青島科技大學(xué)》2017年碩士論文

【摘要】：近年來(lái)熒光納米材料特別是熒光金納米簇已成為國(guó)內(nèi)外科研工作者研究的熱點(diǎn)。熒光金納米簇作為一種簡(jiǎn)單易制備、水分散性好、熒光信號(hào)強(qiáng)、斯托克位移大、生物相容性良好的納米材料已被廣泛應(yīng)用于生物傳感及生物醫(yī)學(xué)等領(lǐng)域。本論文采用較溫和的方法合成熒光金納米簇,以此構(gòu)建新型生物傳感器,并將其應(yīng)用于疾病小分子、腫瘤標(biāo)志物的檢測(cè),成為臨床診斷及治療的一種輔助手段。主要工作概括如下:(1)使用球蛋白包覆合成了一種新型的、穩(wěn)定的熒光金納米簇,并利用它實(shí)現(xiàn)了對(duì)無(wú)機(jī)焦磷酸鹽以及無(wú)機(jī)焦磷酸酶的快速靈敏檢測(cè)。其檢測(cè)原理是利用Cu2+可以猝滅金納米簇?zé)晒獾奶匦?設(shè)計(jì)構(gòu)筑多重?zé)晒鈾z測(cè)生物分子的體系。首先向Cu2+/Au NCs熒光猝滅體系中加入無(wú)機(jī)焦磷酸鹽(PPi),由于PPi可以與Cu2+發(fā)生強(qiáng)絡(luò)合作用,可使猝滅的金納米簇?zé)晒饣謴?fù),實(shí)現(xiàn)了對(duì)生物分子PPi的定量檢測(cè),線性檢測(cè)范圍是0.05-218.125μM,檢測(cè)限為0.02μM。向該混合體系中加入無(wú)機(jī)焦磷酸酶(PPase),可將無(wú)機(jī)焦磷酸鹽PPi水解為磷酸分子,Cu2+重被釋放,納米簇的熒光又被猝滅,從而實(shí)現(xiàn)對(duì)PPase的定量檢測(cè)。實(shí)驗(yàn)結(jié)果表明,該檢測(cè)方法的線性范圍為0.1-8 mU,檢測(cè)限為0.04 mU,并且選擇性較好,有望應(yīng)用于實(shí)際檢測(cè)。(2)利用單一激發(fā)雙發(fā)射金納米簇設(shè)計(jì)了一種新型“關(guān)開(kāi)”式FRET傳感器用于同時(shí)檢測(cè)多種腫瘤標(biāo)志物—甲胎蛋白(AFP)、癌胚抗原(CEA)。AFP適配體修飾的綠色熒光金納米簇(510 nm)以及CEA適配體修飾的紅色熒光金納米簇(650 nm)作為能量供體,二硫化鉬(MoS2)作為能量受體,構(gòu)建新型FRET傳感器,加入目標(biāo)CEA和AFP后記錄510 nm和650 nm處的熒光強(qiáng)度變化同時(shí)定量檢測(cè)這兩種腫瘤標(biāo)志物。AFP線性檢測(cè)范圍為0.5到60 ng/m L,CEA的線性檢測(cè)范圍0.5到120 ng/mL檢測(cè)限(3σ)分別為0.16 ng/m L、0.21 ng/m L。此外該生物傳感器不僅可以實(shí)現(xiàn)準(zhǔn)確定量測(cè)定多種腫瘤標(biāo)志物也可用于可視化的半定量檢測(cè)。更重要的是激光共聚焦熒光顯微鏡實(shí)驗(yàn)證明該生物傳感器可以定性的區(qū)分健康與肝癌病人的血清,此實(shí)驗(yàn)方法具有臨床診斷試驗(yàn)的潛力。
[Abstract]:In recent years, fluorescent nanomaterials, especially fluorescent gold nanoclusters, have become the focus of research at home and abroad. As a kind of simple and easy preparation, gold nanoclusters with good water dispersion, strong fluorescence signal, large Stoke shift and good biocompatibility have been widely used in biosensor and biomedical fields. In this paper, fluorescent gold nanoclusters were synthesized by a mild method, and a new biosensor was constructed. The biosensor was applied to the detection of disease small molecules and tumor markers, and became an auxiliary method for clinical diagnosis and treatment. The main work is summarized as follows: (1) A novel stable fluorescent gold nanocluster was synthesized by globulin coating, and the rapid sensitive detection of inorganic pyrophosphate and inorganic pyrophosphatase was achieved by using it. The detection principle is to design and construct a multiplex fluorescence detection system for biomolecules by using Cu _ 2 to quench the fluorescence characteristics of gold nanoclusters. Firstly, the inorganic pyrophosphate PPian was added to the fluorescence quenching system of Cu2 / au NCs. The fluorescence of the quenched gold nanoclusters could be restored due to the strong complexation of PPi with Cu2, and the quantitative detection of PPi was realized. The linear detection range was 0.05-218.125 渭 m, and the detection limit was 0.02 渭 M. When the inorganic pyrophosphatase PPaseau was added to the mixed system, the inorganic pyrophosphate PPi could be hydrolyzed into phosphoric acid molecule, Cu2, which was rereleased, and the fluorescence of the nanoclusters was quenched, thus the quantitative detection of PPase could be realized. The experimental results show that the linear range of the method is 0.1-8 mU, the detection limit is 0.04 mU, and the selectivity is good. A novel "closed" fret sensor has been designed for simultaneous detection of a variety of tumor markers, such as alpha fetoprotein (AFP), carcinoembryonic antigen (CEA) CEA, AFP aptamer modified green. Color fluorescent gold nanoclusters (510 nm) and CEA aptamer modified red fluorescent gold nanoclusters (650 nm) were used as energy donors. Molybdenum disulfide (MoS _ 2) as an energy receptor, a novel fret sensor was constructed. The fluorescence intensity changes at 510nm and 650nm were recorded after the addition of target CEA and AFP. The linear detection range was 0.5-60 ng/m / mL. The linear detection limit of CEA was 0.16 ng/m / L 0.21 ng/m / mL, respectively. In addition, the biosensor can not only accurately and quantitatively determine many tumor markers, but also can be used for visualized semi-quantitative detection. More importantly, laser confocal fluorescence microscopy has proved that the biosensor can qualitatively distinguish the serum of healthy and liver cancer patients. This experimental method has the potential of clinical diagnosis.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：O657.3

【參考文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 袁智勤;聚乙烯亞胺輔助的熒光金、銀納米材料的合成及應(yīng)用研究[D];湖南大學(xué);2013年

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本文編號(hào)：2021279