双配体功能化荧光金纳米簇的合成及其检测应用研究

发布时间：2018-03-06 13:33

本文选题：双配体功能化金纳米簇　切入点：谷胱甘肽　出处：《青岛科技大学》2017年硕士论文　论文类型：学位论文

【摘要】：作为新兴的荧光探针,双配体功能化的金纳米簇(Au NCs)由于其具有独特的光学物理性质,良好的生物相容性、尺寸比较小、灵敏性高、选择性好等一系列优点,而被广泛应用于生物传感和生物成像中,由于其潜在的应用,蛋白质、聚合物、烷烃硫醇以及多肽等材料被作为配体用于荧光金纳米簇的合成。由于等离子体增强荧光有着荧光信号放大和有效提高检测灵敏度的能力,成为一种新兴技术,新型的核壳金纳米棒结构作为近红外荧光信号打开的传感器,可用于高灵敏,高选择性的检测。本论文主要研究内容如下:(1)在本工作中,我们开发了一种简便的一步策略快速制备双配体功能化的荧光金纳米团簇(Au NCs),并建立了一种“turn on”方法,用于在水溶液和活体细胞中快速和选择性的检测谷胱甘肽(GSH)。所制备的Au NCs显示橙色红色荧光(λem=608 nm),荧光寿命长(5.62μs),斯托克斯位移大(300 nm),稳定性比较好,体系的表征主要通过使用荧光光谱表征、高分辨透射电镜(HRTEM)、动态光散射(DLS)、X射线光电子能谱(XPS)、荧光寿命、红外光谱(IR)、紫外吸收光谱表征。基于Au NCs和GSH之间与Cu2+的竞争性配位诱导的荧光恢复,目前的“turn on”方法对GSH检测提供了高灵敏度和优异的选择性,检测限为9.7 nM。更重要的是,这种“turn on”方法也可以成功应用于可视化和监测Hep G2细胞中GSH水平的变化,为诊断应用提供潜在的应用价值。(2)光学可交叉响应传感器阵列最近被公认为高通量蛋白质检测和分析的强大技术。基于六种近红外荧光双配体功能化金纳米簇(不同种类氨基酸功能化)作为传感受体,开发了能够分析各种蛋白质并区分不同阶段乳腺癌患者血清的多维传感器阵列。这种“化学鼻子/舌头”传感器阵列可同时有效地区分十种蛋白质。线性判别分析(LDA)的响应模式显示出分析物在低至10 nM的浓度下成功分化,识别精度高。等温滴定量热法(ITC)实验表明,Au NCs主要通过氢键和范德华力与蛋白质相互作用。此外,通过成功区分来自乳腺癌患者(早,中,晚)和健康人的血清,证明了该传感器阵列的最大亮点,表明在临床辅助诊断中的潜力很大。(3)本工作中合成了金纳米棒,对金纳米棒进行了高分辨透射电镜和紫外吸收光谱表征,实验结果说明了合成的金纳米棒均匀性比较好,又对金纳米棒进行了二氧化硅的包覆,高分辨透射电镜表征显示了包覆的均匀性。该二氧化硅包覆的金纳米棒将有望用于发展基于等离子体增强荧光的检测体系。
[Abstract]:As a new fluorescent probe, double ligand functionalized gold nanoclusters (au NCs) have a series of advantages, such as unique optical physical properties, good biocompatibility, small size, high sensitivity, good selectivity and so on. And has been widely used in biosensor and imaging, because of its potential applications, proteins, polymers, Alkyl mercaptan and polypeptides are used as ligands in the synthesis of fluorescent gold nanoclusters. Plasma enhanced fluorescence has become a new technology because of its ability to amplify fluorescence signals and improve detection sensitivity. A new structure of core shell gold nanorods, which is opened by near infrared fluorescence signal, can be used to detect high sensitivity and selectivity. The main contents of this thesis are as follows: 1) in this work, We have developed a simple, one-step strategy for the rapid preparation of double ligand functionalized gold nanoclusters, au NCsN, and a "turn on" method has been established. Au NCs was prepared for rapid and selective detection of glutathione glutathione (GSH) in aqueous solution and in vivo cells. The au NCs showed orange red fluorescence (位 em=608 NmM), long fluorescence lifetime of 5.62 渭 s-1, and a large Stokes shift of 300nmG, and showed good stability. The system was characterized by fluorescence spectroscopy, high resolution transmission electron microscopy (TEM), dynamic light scattering (DLSX) X-ray photoelectron spectroscopy (XPS), and fluorescence lifetime. Based on the competitive coordination between au NCs and GSH and Cu2 induced fluorescence recovery, the current "turn on" method provides high sensitivity and excellent selectivity for GSH detection with a detection limit of 9.7 nm. This "turn on" method can also be successfully used to visualize and monitor the changes of GSH levels in Hep G2 cells. Providing potential application value for diagnostic applications. Optical cross responsive sensor arrays have recently been recognized as a powerful technique for high throughput protein detection and analysis. Based on six near infrared fluorescent double ligand functionalized gold nanoclusters (NIR). Different kinds of amino acid functionalization) as sensing receptors, A multi-dimensional sensor array has been developed that can analyze various proteins and distinguish the serum of breast cancer patients at different stages. This "chemical nose / tongue" sensor array can effectively distinguish ten proteins at the same time. The response patterns of LDAs showed that the analytes differentiated successfully at concentrations as low as 10nM. Isothermal drop quantitative calorimetry (ITC) test showed that au NCs interacted with protein mainly through hydrogen bond and van der Waals force. In addition, by successfully distinguishing serum from breast cancer patients (early, middle, late) and healthy people, The results show that gold nanorods have been synthesized in this work and characterized by high resolution transmission electron microscopy (TEM) and UV absorption spectra. The experimental results show that the synthesized gold nanorods have good uniformity, and the gold nanorods are coated with silica. The high resolution transmission electron microscopy (TEM) shows the uniformity of the coating. The silica coated gold nanorods are expected to be used in the development of plasma-enhanced fluorescence detection systems.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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