An immunoassay using antibody-gold nanoparticle conjugate, silver enhancement and flatbed scanner
An immunoassay using antibody-gold nanoparticle conjugate, silver enhancement and flatbed scanner
复制标题
DOI：
10.1007/s10404-008-0298-0
复制
发表时间：
2009-01-01
影响因子：
2.8
通讯作者：
Lin, Yu-Cheng
中科院分区：
文献类型：
--
作者：
Yeh, Chia-Hsien;Hung, Ching-Yuan;Lin, Yu-Cheng
本研究报告了一种方便的免疫分析方法，使用抗体-金纳米颗粒（Ab-AuNP）结合物作为报告分子和平板扫描仪进行光学扫描和测量的免疫反应信号。引入了银增强反应，即一种信号放大方法，其中银离子被还原为银金属，以放大检测信号。本研究的整个框架基于（1）直接免疫测定（双层形式），其中抗原直接固定在芯片上，以及（2）夹心免疫测定[三层形式，过程类似于酶联免疫吸附测定（ELISA）]，其中包含一抗、二抗（抗体-纳米颗粒缀合物）和抗原。实验数据表明，微尺度银沉淀现象是由Ab-AuNP缀合物催化的。即使在5 pM的IgG-AuNP浓度下，也可以通过肉眼观察到该反应。讨论了样品浓度与检测信号之间的关系，样品抗原的检测限（夹心法）为1 ng/mL。使用平板扫描仪，Ab-AuNP缀合物和银增强反应，构建了一种新的免疫分析。
This study reports a convenient immunoassay using antibody-gold nanoparticle (Ab-AuNP) conjugate as a reporter molecule and a flatbed scanner for the optical scanning and measuring of the immuno-reaction signal. The silver enhancement reaction, a signal amplification method in which silver ions are reduced to silver metal, is introduced to magnify the detection signal. The whole framework of the study is based on (1) the direct immunoassay (two-layer format) in which the antigen is directly immobilized on the chip, and (2) the sandwich immunoassay [three-layer format, process similar to enzyme-linked immunosorbent assay (ELISA)], which contains a primary antibody, a secondary antibody (antibody-nanoparticle conjugate) and antigens. The experimental data show that the micro-scale silver precipitation phenomenon is catalyzed by Ab-AuNP conjugates. This reaction can be observed by the naked eye, even at an IgG-AuNPs concentration of 5 pM. The relationships between sample concentration and detection signal are discussed, and the detection limit (sandwich assay) for the sample antigen is 1 ng/mL. Using a flatbed scanner, Ab-AuNP conjugates and a silver enhancement reaction, a new immunoassay is constructed.