东篱科研大数据发现系统（DRDS）

位置：成果数据库 > 期刊 > 期刊详情页

基于近红外上转换荧光共振能量传递体系的均相免疫分析

ISSN号：1000-0593
期刊名称：《光谱学与光谱分析》
时间：0
分类：O472.3[理学—半导体物理;理学—物理]
作者机构：[1]中国科学院长春光学精密机械与物理研究所激发态重点实验室,吉林长春130033, [2]中国科学院研究生院,北京100049
相关基金：国家自然科学基金项目（60771051 10674132 20603035 10904142）资助

关键词：近红外, 上转换, 荧光共振能量传递, Near infrared, Upconversion, Fluorescence resonance energy transfer

中文摘要：

在上转换纳米晶（UCNPs）为供体的荧光共振能量传递（FRET）生物均相检测体系中,弱的供体光强度使FRET信号难于检测,同时还有来自生物基质的自发荧光干扰。这使得UCNPs不产生背景荧光和散射光的优点不能够充分地体现。针对这个问题,作者利用在800nm处有强近红外光的NaYF4：Yb3＋,Tm3＋UCNPs作为供体,在784nm处有表面等离子共振吸收带的金纳米粒子（GNPs）作为受体构建了新型的FRET体系。UCNPs偶联抗体（goat antihumanIgG）及GNPs偶联抗原（humanIgG）,在抗原抗体免疫亲和作用下两者距离靠近;UCNPs荧光光谱和GNPs吸收光谱有效交叠,使FRET发生。当体系中加入单纯humanIgG,竞争性地争夺与goatantihumanIgG结合位点,破坏FRET构建,供体近红外光增强。根据此对应关系,确定humanIgG检测限为5μg·mL-1。这种方法可适用于更广泛的荧光分析。

英文摘要：

A FRET based assay utilizing NaYF4：Yb3＋,Tm3＋ UCNPs as an energy donor,which can emit intense near infrared（NIR） upconversion emission around 800 nm ranges under illumination with a 980 nm laser,and GNPs as an energy acceptor,which has an surface plasmon absorption maximum at 784 nm,was demonstrated.Their optical properties satisfy the requirement of spectral overlap between donors and acceptors for FRET.A model assay for human IgG was then constructed,in which amino-functionalized NaYF4：Yb3＋,Tm3＋ UCNPs and GNPs were first prepared and then conjugated with the antibody（goat antihuman IgG） and antigen（human IgG）,respectively.The mutual affinity of the antigen and antibody brought the nanocrystals close enough together to allow the FRET to occur,resulting in a significant quenching of UCNPs upconversion emission at 800 nm.When free human IgG was added to the immunocomplex,it competitively binds to UCNPs-goat antihuman IgG,thereby replacing human IgG-GNPs from the immunocomplex and inhibits the FRET process.As a result,the gradually increasing the NIR emission was observed.The authors associate the fluorescence enhancement effect with the concentration of human IgG.Under our experimental conditions,the detection limit is 5 μg·mL-1.This approach is expected to be extended to the detection of other biological fields,enabling measurements without background fluorescence interference.