基于金磁微粒的即时检测用免疫探针的构建及微结构调控

项目名称： 基于金磁微粒的即时检测用免疫探针的构建及微结构调控

项目编号： No.21505089

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 杨冬

作者单位： 陕西科技大学

项目金额： 21万元

中文摘要： 针对即时检测的高灵敏度、高特异性及定量的需求，本研究以兼具光学、磁学双重特性的核壳Fe304/Au纳米粒为载体，以氨基末端脑尿肽前体（NT-proBNP）为检测目标，制备微结构可控的金磁免疫探针，建立简单、经济、快速的免疫层析定量检测系统。首先，通过多种表面改性方法，获得具备不同表面特性的单分散性的Fe304/Au纳米粒；再通过“纳米-生物界面控制”偶联单抗NT-proBNP，制备微结构可控的金磁免疫探针；最后在免疫层析系统实现待检物的磁定量检测。本课题在研究Fe304/Au纳米粒的稳定性及机理的基础上，揭示Fe304/Au纳米粒的表面特性对金磁免疫探针微结构调控的影响规律，并建立探针微结构形成的热力学及动力学方程；揭示“结构-检测性能”关系规律，量化“结构-磁信号强度”的内在联系，建立POCT免疫定量检测模型，为基于金磁纳米微粒的即时检测及体外诊断试剂的设计与开发提供新方法及理论支持。

中文关键词： 金磁免疫探针；金磁纳米微粒；即时检测；微结构调控

英文摘要： The major target of point-of-care testing (POCT) is to get higher sensitivity, specifity and reach quantitative level. Composite Fe3O4/Au nanoparticle have arracted considerable interest in medicine due to their unique physical and chemical properties, including optical properties, superparamagnetic properties and flexibility to biofunctionalization for downstream application. In this study, we explore Fe3O4/Au nanoparticle as carriers, and simultaneously cardiac marker N-terminal pro-B-type natriuretic peptide (NT-proBNP) as model to realize simple, inexpensive, sensitive, quantitative lateral flow immunoassay. Firstly, Fe3O4/Au nanoparticles with different surface properties and mono-dispersion are fabricated by harnessing the surfactant, ligand and polymer synergistic cooperation. Secondly, the immunoassay probes with regulated micro-structure can be synthesized by conjugating monoclonal antibodies (NT-proBNP) to the surface of gold magnetic nanoparticle. At last, the probes are used in lateral flow immunoassay for NT-proBNP quantitative detection. In the study, we investigate the stability mechanism of composite Fe3O4/Au nanoparticles before and after surface modification, reveal how surface characteristics and composition effect on the regulation of micro-structure of the immunoassay probes in detail, establish thermodynamic and kinetic equations for the formation of probes’ micro-structure, investigate the relationship between micro-structure and detection results, including sensitivity, specificity and magnetic signal intensity. The study will not be of benefit to POCT detection based on gold magnetic nanoparticles, but also the design and development of native vitro diagnostic reagents.

英文关键词： Gold magnetic immunoassay probe;Gold magnetic nanoparticle;point-of-care testing;micor-structure regulation