基于纳米材料荧光检测生物分子的研究

发布时间：2018-01-13 15:35

本文关键词：基于纳米材料荧光检测生物分子的研究　出处：《湘潭大学》2015年硕士论文　论文类型：学位论文

【摘要】：纳米金和氧化石墨烯作为目前新兴的两种重要的纳米材料,在实际生活中各个领域的应用越来越广泛,特别是在生命科学、药物分析中的应用。两种纳米材料均有着自身优良的特性及作为纳米材料的特性,为生物传感器的构建提供条件。核酸适配体对靶物质的高特异识别能力和高亲和力,为检测物质消除干扰,且可进行功能化修饰在纳米金或者氧化石墨烯上用来制备核酸适配体传感器。荧光探针是指利用荧光信号与目标物的浓度存在关系,通过观察荧光强度的变化来检测靶物质。本论文通过构建不同机理的荧光探针检测生物小分子物质包括腺苷、多巴胺以及卡那霉素。由此方法构建的荧光探针检测简便,具备高灵敏度、高选择性。(1)第一章介绍了以下内容:纳米金和氧化石墨烯的性质特征、制备方法以及应用研究;核酸适配体的制备及应用;荧光传感器的机理分析;本论文的研究意义及主要研究内容。(2)基于纳米金和氧化石墨烯构建的荧光探针检测溶液中的分析物质。在本实验中,腺苷适配体可通过Au-S键修饰在纳米金表面,无靶物质腺苷时,纳米金-适配体可通过π-π堆叠作用吸附在氧化石墨烯表面,纳米金-适配体-氧化石墨烯的复合物沉淀将会生成;在靶物质腺苷存在时,纳米金-适配体会存在于上清液中,不会全部与氧化石墨烯形成沉淀。加入荧光素,无靶物质腺苷的上清液荧光较强,有靶物质腺苷的上清液荧光较弱。通过检测荧光的强度,不同浓度的腺苷可被检测。线性范围5?10-8 M到5?10-7 M,检出限为5.8?10-9 M。这个荧光探针对腺苷的检测特异性强,其他类似物均无明显干扰。并且此荧光探针对于实际样品中的腺苷检测具有重要的意义。(3)基于氨基芘和氧化石墨烯构建的荧光探针检测多巴胺。在无多巴胺存在时,荧光物质氨基芘的荧光可被荧光淬灭剂氧化石墨烯淬灭,主要是通过氨基芘与氧化石墨烯之间的?-?堆叠作用。在多巴胺存在时,多巴胺将会吸附在氧化石墨烯的表面,使氨基芘从其表面析出。由于多巴胺与氧化石墨烯之间的强吸附作用,这个荧光探针具有较好的灵敏度和选择性。线性范围为4×10-9 M to 4×10-7 M,检出限为7.53 nM。本实验探讨了氧化石墨烯与适配体对多巴胺的吸附作用,手段为荧光光谱、红外光谱、以及朗缪尔吸附理论分析。可知氧化石墨烯对多巴胺有更强的吸附作用,因此两者的强吸附作用使该荧光探针可对样品中多巴胺进行检测。(4)基于阳离子体荧光产物、适配体、氧化石墨烯及DNA酶的相互作用检测靶物质卡那霉素。阳离子体荧光产物在加入适配体时荧光淬灭,加入氧化石墨烯,荧光进一步淬灭,当加入靶物质卡那霉素和DNA酶时,带有阳离子荧光物质的适配体从氧化石墨烯表面析出,在DNA酶的催化作用下,适配体降解,释放靶物质,进入循环,荧光增强并放大。该荧光探针的线性范围为2×10-8 M到10×10-8 M,检出限为8.67 nM,且实验无明显干扰,可检测实际样品中的卡那霉素。
[Abstract]:Gold nanoparticles and graphene oxide as two important nanomaterials are currently emerging in various fields, more and more widely in real life, especially in life science, application in pharmaceutical analysis. Two kinds of nano materials has its excellent properties and as nanometer material properties, provide the conditions for the construction of biological sensors. High specific recognition ability of the aptamer target material and high affinity for the detection of substances eliminating interference, and can be used for the preparation of functionalized aptamer sensor in gold nanoparticles or graphene oxide. The fluorescent probe refers to the concentration by fluorescence signal and the presence of a target, by observing the changes the fluorescence intensity of the detected target material. This paper through the material detection of biological small molecule fluorescent probe to construct different mechanisms including adenosine, dopamine and kanamycin. The method. The fluorescent probe to detect the construction is simple, with high sensitivity, high selectivity (1). The first chapter introduces the following contents: the properties of gold nanoparticles and graphene oxide, research methods and application of preparation; preparation and application of aptamer fluorescence sensor; mechanism analysis; the main research contents and research significance of this thesis. (2) analysis of matter fluorescence probe detection solution of gold nanoparticles and graphene oxide based. In this experiment, adenosine aptamer can be modified on the surface of gold nanoparticles through the Au-S key, no target adenosine, gold nanoparticle aptamer through Pi Pi stacking adsorbed on the surface of oxidation graphene composite gold nanoparticles aptamer - graphene oxide precipitation will be generated in the presence of target material; adenosine, nano gold adaptation experience existed in the supernatant, and not all of the graphene oxide precipitate. With fluorescein, no target material The supernatant of adenosine strong fluorescence, a weak fluorescence supernatant of target material adenosine. Detected by fluorescence intensity, different concentrations of adenosine can be detected. The linear range of 5 to 5? 10-8 M? 10-7 M, the detection limit was 5.8? 10-9 M. the fluorescence probe for adenosine detection specificity, there were no other analogues significant interference. And the fluorescence probe has important significance for adenosine in actual sample testing. (3) the fluorescent probe to detect dopamine aminopyrene and graphene oxide based. In the absence of dopamine exists, fluorescent substances can be aminopyrene fluorescence quencher graphene oxide is mainly through quenching. Aminopyrene and graphene oxide?? stacking. In the presence of dopamine, dopamine will be adsorbed on the surface of graphene oxide, the amino pyrene from the surface precipitation. Due to the strong adsorption between dopamine and graphene oxide as With this, fluorescence probe has a good sensitivity and selectivity. The linear range of 4 * 10-9 4 * to M 10-7 M, the detection limit was 7.53 nM. in this study, the adsorption of graphene oxide and aptamer of dopamine, means of fluorescence spectroscopy, infrared spectroscopy, and Langmuir adsorption theory. The graphite oxide graphene has a strong adsorption of dopamine, so the strong adsorption of both the fluorescent probe to detect samples. Dopamine (4) cationic body fluorescent products, based on aptamer, interaction detection of graphene oxide and DNA enzyme target material of kanamycin. Cationic fluorescent products in fluorescent body aptamer when quenching, adding graphene oxide, fluorescence quenching further, when adding the target substance of kanamycin and DNA enzyme, a cationic fluorescent substance aptamer from the surface of graphene oxide precipitates in DNA enzyme catalyzed. The target material was degraded and released into the loop under the condition of aptamer degradation. The fluorescence enhancement and amplification were achieved. The linear range of the fluorescence probe is 2 * 10-8 M to 10 * 10-8 M, the detection limit is 8.67 nM, and the experiment has no obvious interference. It can detect kanamycin in the actual sample.

【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：O657.3;TB383.1

【共引文献】