纳米银拉曼增强基底的制备及其在蛋白质检测中的应用

发布时间：2018-02-17 00:21

本文关键词： SERS 蛋白质自组装技术免标记　出处：《南京理工大学》2017年硕士论文　论文类型：学位论文

【摘要】：表面增强拉曼光谱(SERS)作为指纹识别和无损检测技术在生物分子检测领域表现了其他检测手段无法比拟的优势:痕量、无损、快速、指纹式识别,能够提供丰富的生物分子结构信息及其振动信息。本文采用自组装的方法分别制备了可用于蛋白质分子快速检测的固态SERS基底和有望用于生物活体中蛋白检测的悬浮态SERS基底。并将其成功应用到蛋白质检测的应用中。主要研究内容如下:(1)利用不同的方法合成银纳米材料,重点研究了 LeeMeisel的水热合成法,并对其合成工艺做了改进。通过控制前期沸腾的时间,成功地合成出了尺寸均一的类球形的银纳米颗粒,其粒径在50-70nm;(2)优化了传统的二次氧化法,通过加入过量的浓硫酸及调整石墨粉和高锰酸钾的加料速度等,制备了分散性更好的氧化石墨烯。并通过TEM、UV-Vis及拉曼仪等对其进行了表征。(3)利用自组装技术在玻璃片表面组装银纳米颗粒。通过控制时间在玻璃表面形成具有良好的表面增强拉曼效应的"热点",并探究了该基底材料对不同的酸化蛋白以及不同蛋白浓度的表面增强拉曼的增强效果,同时也使用了扫描电镜等对该基底材料的形貌表征,其检测极限达到了 0.1μ g/mL。(4)通过引入悬浮性良好的氧化石墨烯作为平台,通过调节NaCl、HCl的加入量来打开氧化石墨烯和银纳米颗粒表面的双电子层,使银纳米颗粒进行可控的团聚并负载到氧化石墨烯上形成有SERS效应的基底材料,从而实现免标记蛋白质的痕量检测和分析,其检测线达到0.4μg/mL。
[Abstract]:Surface-Enhanced Raman Spectroscopy (SERS) as a fingerprint identification and nondestructive testing technology in the field of biomolecular detection has shown unparalleled advantages: trace, non-destructive, fast, fingerprint identification, In this paper, we have prepared solid state SERS substrates for rapid detection of protein molecules and those expected to be used in the detection of proteins in living organisms by using self-assembly method, which can provide abundant information on the structure and vibration of biomolecules. The suspended SERS substrate was successfully applied to protein detection. The main research contents are as follows: 1) the silver nanomaterials were synthesized by different methods. The hydrothermal synthesis method of LeeMeisel has been studied emphatically, and the synthesis process has been improved. By controlling the boiling time in the early stage, the spherical silver nanoparticles of uniform size have been successfully synthesized, the particle size of which is 50-70 nm ~ (2)) and the traditional secondary oxidation method has been optimized. By adding excessive concentrated sulfuric acid and adjusting the feeding speed of graphite powder and potassium permanganate, Graphene oxide with better dispersity was prepared and characterized by TEM UV-Vis and Raman spectrometer. Silver nanoparticles were assembled on glass surface by self-assembly technique. The silver nanoparticles were formed on glass surface by controlling time. The "hot spot" of the surface-enhanced Raman effect and the enhancement effect of the substrate on different acidified proteins and different protein concentrations were investigated. At the same time, scanning electron microscopy was used to characterize the morphology of the substrate. The detection limit reached 0.1 渭 g / m 路L ~ (4). Graphene oxide with good suspension was introduced as the platform. The double electron layer on the surface of graphene oxide and silver nanoparticles was opened by adjusting the amount of NaCl-HCl, which allowed the controlled agglomeration of silver nanoparticles and the loading of silver nanoparticles onto graphene oxide to form a substrate material with SERS effect. Thus, the trace detection and analysis of non-labeled protein were realized, and the detection line reached 0.4 渭 g / mL.
【学位授予单位】：南京理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TB383.1;Q6-33

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