基于核酸适配体与金纳米粒子探针检测凝血酶的共振光散射分析

发布时间：2018-04-04 11:34

本文选题：金纳米粒子　切入点：核酸适配体　出处：《河北大学》2008年硕士论文

【摘要】： 蛋白质是构成生物体的重要组成部分之一,是一切生命的物质基础,承载着生物体完成各项生物功能的任务。因此,蛋白质的分析在生命科学的研究中具有重要的意义,广泛应用于生物学、医学诊断等领域。传统的蛋白质检测往往需要荧光标记、凝胶电泳和放射性自显影等繁杂操作,在一定程度上限制了人们对这些生命物质的深入研究。随着生命科学的发展,如何对蛋白质进行快速、简便、高效分析,已经成为研究的热点问题。本文以凝血酶为研究模型,利用核酸适配体(aptamer)与金纳米粒子(gold nanoparticle)技术相结合制作生物传感器,以核酸适配体作为蛋白质识别元件,发展了蛋白质分析的新方法,该方法快速、灵敏、操作简单而且具有很高的选择性。本文分为两个部分：第一部分：基于核酸适配体标记的金纳米粒子探针测定凝血酶的共振光散射分析。核酸适配体是一种具有特殊功能的寡聚核苷酸,它能与目标分子,包括核酸、蛋白质、药物等特异性结合,从而实现目标分子的测定。以标记核酸适配体的金纳米粒子为探针,通过核酸适配体与凝血酶的特异性作用,金纳米粒子发生自组装,形成网状结构,导致共振光散射信号的增强,从而定量测定了a凝血酶。该方法操作简便,具有较高的灵敏度,为蛋白质的检测开辟了一条新途径。第二部分：基于匹配DNA链标记的金纳米粒子探针与核酸适配体作用测定凝血酶的共振光散射分析。本文设计了两条与核酸适配体匹配的寡核苷酸链并将其分别标记到金纳米粒子上,当金纳米粒子探针与核酸适配体混合,此探针与核酸适配体的杂交导致金纳米粒子的聚集并引起共振光散射的强烈增强；而核酸适配体与凝血酶的特异性结合将抑制此探针与核酸适配体的杂交并导致共振光散射强度的降低,降低的程度与凝血酶的浓度有关,从而可以定量测定凝血酶。该方法只要改变核酸适配体和相应的探针序列,即可有望用于其它蛋白质的分析,为蛋白质的检测提供了新的思路和手段。
[Abstract]:Protein is one of the important components of organism and the material basis of all life.Therefore, protein analysis plays an important role in life science research and is widely used in biology, medical diagnosis and other fields.Traditional protein detection often requires a lot of operations such as fluorescent labeling, gel electrophoresis and radioautography, which to some extent limits the further study of these living substances.With the development of life science, how to analyze proteins quickly, easily and efficiently has become a hot issue.In this paper, a novel method for protein analysis was developed by using thrombin as a research model, using aptamer (a nucleic acid aptamer) and gold nanoparticles (gold nanoparticles articlein) technology to produce biosensor, using aptamer of nucleic acid as a protein recognition element.It is sensitive, simple and selective.This paper is divided into two parts:Part I: resonance light scattering (RLS) analysis of thrombin by gold nanoparticles probe labeled with aptamer of nucleic acid.Nucleic acid aptamer is a kind of oligonucleotide with special function. It can bind to target molecule, including nucleic acid, protein, drug and so on, so as to realize the determination of target molecule.Gold nanoparticles labeled with nucleic acid aptamers were used as probes. Through the specific interaction between aptamers and thrombin, gold nanoparticles were self-assembled and formed a network structure, which led to the enhancement of resonance light scattering (RLS) signals.Thus, a thrombin was quantitatively determined.The method is easy to operate and has high sensitivity, which opens a new way for protein detection.Part two: resonance light scattering (RLS) analysis of thrombin based on the interaction between gold nanoparticles labeled with DNA chain and aptamer of nucleic acid.Two oligonucleotide chains matching the aptamer of nucleic acid were designed and labeled onto gold nanoparticles respectively when the gold nanoparticles probe was mixed with the aptamer of nucleic acid.The hybridization of the probe with the aptamer of nucleic acid leads to the aggregation of gold nanoparticles and the enhancement of resonance light scattering.The specific binding of aptamer and thrombin will inhibit the hybridization between the probe and the aptamer of nucleic acid and lead to the decrease of the intensity of resonance light scattering (RLS), which is related to the concentration of thrombin, so the thrombin can be quantitatively determined.This method can be used in the analysis of other proteins by changing the aptamer of nucleic acid and the corresponding probe sequence, which provides a new way for protein detection.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R346

【引证文献】