Flotation Immunoassay: Masking the Signal from Free Reporters in Sandwich Immunoassays

Flotation Immunoassay: Masking the Signal from Free Reporters in Sandwich Immunoassays
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DOI:
10.1038/srep24297
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发表时间:
2016-04-14
期刊:
影响因子:
4.6
通讯作者:
Willson, Richard C.
Willson, Richard C.
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Chen, Hui;Hagstrom, Anna E. V.;Willson, Richard C.

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在这项工作中,我们证明了信号掩蔽试剂与适当的捕获抗体载体一起可以消除夹心免疫测定中的洗涤步骤。以辣根过氧化物酶化学发光为报告系统,亮蓝FCF为信号掩蔽剂,漂浮的二氧化硅微泡为捕获抗体载体,建立了漂浮免疫分析平台。只有捕获在微泡上的报告分子漂浮在染料上方,并以分析物依赖性方式变得可见。这些FI能够检测低至阿托摩尔水平的蛋白质和少至106个病毒颗粒。这种信号掩蔽策略代表了一种在实验室和床旁环境中进行简单、灵敏和定量免疫测定的新方法。
In this work, we demonstrate that signal-masking reagents together with appropriate capture antibody carriers can eliminate the washing steps in sandwich immunoassays. A flotation immunoassay (FI) platform was developed with horseradish peroxidase chemiluminescence as the reporter system, the dye Brilliant Blue FCF as the signal-masking reagent, and buoyant silica micro-bubbles as the capture antibody carriers. Only reporters captured on micro-bubbles float above the dye and become visible in an analyte-dependent manner. These FIs are capable of detecting proteins down to attomole levels and as few as 106 virus particles. This signal-masking strategy represents a novel approach to simple, sensitive and quantitative immunoassays in both laboratory and point-of-care settings.