【摘要】：自上世紀(jì)90年代以來,納米技術(shù)獲得了持續(xù)發(fā)展。由于其獨(dú)特的物理和化學(xué)性質(zhì),納米材料得到了廣泛的研究和應(yīng)用。新型納米材料與熒光、電化學(xué)技術(shù)的結(jié)合,為分析化學(xué)帶來了更有效的分析方法。細(xì)胞色素C是細(xì)胞凋亡的重要信息物質(zhì)之一。發(fā)展簡單、快速的細(xì)胞色素C的檢測方法對于臨床檢測和生物學(xué)研究具有重要意義。在本論文中,研究了將納米技術(shù)與光電化學(xué)、熒光分析技術(shù)相結(jié)合,開發(fā)細(xì)胞色素C的快速檢測方法。具體包括如下三個(gè)方面的工作：(1)CdS/ZnIn2S4/TiO2三維異質(zhì)結(jié)構(gòu)的制備及其光電性能的研究。以Ti02納米管陣列為基底,通過簡單水熱法和連續(xù)離子層吸附和反應(yīng)法,制備三維CdS/ZnIn2S4/TiO2異質(zhì)結(jié)構(gòu)。在AM1.5G輻射下,CdS/ZnIn2S4/TiO2顯示了優(yōu)良的光電性能,光電流達(dá)到4.3 mA/cm2,光電轉(zhuǎn)換效率為2%。本研究為更有效地利用太陽光光譜提供了一種新型三維異質(zhì)結(jié)構(gòu)體系的設(shè)計(jì)路線。與此同時(shí),本研究對材料進(jìn)行了光催化降解性能測試,值得注意的是,在持續(xù)90 min的光照后,100%的二氯苯氧基乙酸(2,4-D)被移除,而使用ZnIn2S4/TiO2和TiO2納米管陣列在同一條件相同時(shí)間內(nèi)分別只能達(dá)到72.3%和39.1%的去除率。穩(wěn)定性實(shí)驗(yàn)證明材料具有良好的化學(xué)穩(wěn)定性。此外,本課題詳細(xì)地研究了該材料進(jìn)行光催化降解的機(jī)制。(2)基于Au/CdS/ZnIn2S4/TiO2復(fù)合電極材料的無標(biāo)記細(xì)胞色素C光電感器的研究。通過恒電位電化學(xué)沉積法將金納米顆粒沉積到CdS/ZnIn2S4/TiO2復(fù)合電極表面,獲得性能優(yōu)異的Au/CdS/ZnIn2S4/TiO2納米復(fù)合材料,并將其作為光電極：以巰基修飾的細(xì)胞色素C特異性適配體為識(shí)別物質(zhì)與電極結(jié)合,構(gòu)建無標(biāo)記光電適配體傳感器實(shí)現(xiàn)對細(xì)胞色素C的定量檢測。檢測原理：通過檢測適配體-細(xì)胞色素C相結(jié)合,導(dǎo)致的光電流變化情況,間接測定細(xì)胞色素C的濃度。本方法中,利用對細(xì)胞色素C具有高特異性識(shí)別性能的適配體和高靈敏電化學(xué)檢測方法的結(jié)合,使傳感體系具備高度的選擇性和相對較低的檢測限(1.0pM)。此外,進(jìn)一步的實(shí)驗(yàn)數(shù)據(jù)證明了該傳感器具有良好的重現(xiàn)性和穩(wěn)定性。(3)基于VS2納米片的新型生物傳感器快速熒光檢測細(xì)胞色素C的研究。由于細(xì)胞色素C本身不具有熒光,所以無法直接通過檢測其自身熒光來進(jìn)行定性或定量。本研究通過間接熒光恢復(fù)的方法對其進(jìn)行測定。通過簡單溶液方法合成具有強(qiáng)熒光猝滅能力的VS2納米片,并將其作為傳感響應(yīng)平臺(tái)；末端用熒光染料羧基熒光素標(biāo)記的能與細(xì)胞色素C特異性結(jié)合的適配體作為識(shí)別探針。VS2納米片能有效地吸附探針并猝滅探針末端熒光團(tuán)的熒光。然而,當(dāng)熒光探針先與細(xì)胞色素C進(jìn)行孵育,再與VS2納米片溶液混合,探針的熒光將不會(huì)被猝滅。由于所用適配體的高特異性能,傳感器對細(xì)胞色素C顯示出了高度的選擇性和靈敏度,其線性范圍為0.75nM-50μM,檢測限為0.5 nM,同一檢測條件下對生物體內(nèi)其他干擾物基本無檢出。
[Abstract]:Nanotechnology has been continuously developing since the 1990s. Due to its unique physical and chemical properties, the nano-materials have been widely studied and applied. The combination of new nano-materials with fluorescence and electrochemical technology has brought more effective analytical methods for analytical chemistry. Cytochrome C is one of the important information materials of cell apoptosis. The development of simple and rapid detection of cytochrome C is of great significance for clinical detection and biological research. In this paper, the method of rapid detection of cytochrome C was developed by combining nano-technology with photochemistry and fluorescence analysis. The method comprises the following three aspects: (1) the preparation of the CdS/ ZnIn2S4/ TiO2 three-dimensional heterostructure and the research on the photoelectric property thereof. Three-dimensional CdS/ ZnIn2S4/ TiO2 heterostructures were prepared by simple hydrothermal method and continuous ion-layer adsorption and reaction. CdS/ ZnIn2S4/ TiO2 showed excellent photoelectric properties under the radiation of AM1.5G, and the photocurrent reached 4. 3mA/ cm2 and the photoelectric conversion efficiency was 2%. The present study provides a new design route of a new three-dimensional heterostructure system for the more efficient use of the solar spectrum. At the same time, the present study tested the photocatalytic degradation of the material, noting that 100% of the dichlorophenoxyacetic acid (2, 4-D) was removed after 90 min of illumination, The use of ZnIn2S4/ TiO2 and TiO2 nanotube arrays can only reach 72.3% and 39.1% of the removal rate in the same time. The stability experiment proves that the material has good chemical stability. In addition, the mechanism of photocatalytic degradation of the material was studied in detail in this paper. (2) The study of the non-labeled cytochrome C photoinductor based on the Au/ CdS/ ZnIn2S4/ TiO2 composite electrode material. the gold nanoparticles are deposited on the surface of the CdS/ ZnIn2S4/ TiO2 composite electrode by a constant potential electrochemical deposition method, and the Au/ CdS/ ZnIn2S4/ TiO2 nano composite material with excellent performance is obtained, and the quantitative detection of the cytochrome C is realized by constructing a non-labeled photoelectric aptamer sensor. the detection principle: the concentration of the cytochrome c is indirectly determined by detecting the change of the photocurrent caused by the combination of the aptamer-cytochrome c. In the method, the sensor system is provided with a high selectivity and a relatively low detection limit (1,0pM) by combining an aptamer with high specificity identification performance and a high-sensitivity electrochemical detection method for cytochrome C. Furthermore, further experimental data demonstrate that the sensor has good reproducibility and stability. (3) Study on the rapid fluorescence detection of cytochrome C by novel biosensor based on VS2 nanosheet. Since the cytochrome C itself does not have fluorescence, it is not possible to make a qualitative or quantitative determination by detecting its own fluorescence. This study was determined by means of indirect fluorescence recovery. The VS2 nanosheet with strong fluorescence quenching capacity is synthesized by a simple solution method, and is used as a sensing response platform, and the terminal is used as an identification probe with a fluorescent dye-based fluorescein-labeled aptamer capable of specifically binding to the cytochrome C. The VS2 nanosheet can effectively adsorb the probe and wipe out the fluorescence of the fluorophore at the end of the probe. However, when the fluorescent probe is first incubated with cytochrome C and then mixed with the VS2 nanosheet solution, the fluorescence of the probe will not be inactivated. Due to the high specific energy of the aptamer used, the sensor showed a high degree of selectivity and sensitivity to the cytochrome C, with a linear range of 0.75nM to 500.mu. M, with a detection limit of 0.5 nM, and the other interfering substances in the organism were substantially free of detection under the same detection conditions.
【學(xué)位授予單位】：湖南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：TB383.1;O657.3
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本文編號(hào)：2336259