One-step in situ solid-substrate-based whole blood immunoassay based on FRET between upconversion and gold nanoparticles
One-step in situ solid-substrate-based whole blood immunoassay based on FRET between upconversion and gold nanoparticles
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基于上转换和金纳米颗粒之间的 FRET 的一步式原位固体基质全血免疫分析
DOI：
10.1016/j.bios.2016.11.003
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发表时间：
2017-06-15
影响因子：
12.6
通讯作者：
Kong, Xianggui
中科院分区：
文献类型：
--
作者：
Li, Cuixia;Zuo, Jing;Kong, Xianggui
尽管它们具有普遍的临床应用，但当前基于荧光的免疫测定法面临着严峻的挑战，例如当前异质方法中先进的血清/血浆分离和繁琐的洗涤过程，以及均相方法中的颗粒聚集、低灵敏度和窄线性范围。本文通过结合传统的荧光共振能量转移（FRET）技术（上转换纳米颗粒（UCNP）和金纳米颗粒（GNP）之间）和基于固体基质的免疫分析技术，在全血样本的新型一步原位免疫分析中解决了这些紧迫问题。山羊 IgG (gIgG) 的检测限低，缓冲液中为 0.042 μg/mL，20 倍稀释全血样品中为 0.51 μg/mL，血液样品中的线性范围为 0.75 μg/mL 至 60 μg/mL。据我们所知，这是第一个基于原位固体基质的全血样本免疫分析，具有较大的线性检测范围。这一进展为直接在全血中对各种生物分子进行快速、灵敏的免疫测定提供了一个有前景的平台，无需繁琐的分离、洗涤步骤和聚集问题。
Despite their general clinical applications, current fluorescence-based immunoassays are confronted with serious challenges, e.g. the advance serum/ plasma separation and the tedious washing process in current heterogeneous approaches, and aggregation of particles, low sensitivity and the narrow linear range in homogeneous approaches. In this paper, these urgent problems were solved in a novel one-step in situ immunoassay of whole blood samples by combining the traditional fluorescence resonance energy transfer (FRET) technology (between upconversion nanoparticles (UCNPs) and gold nanoparticles (GNPs)) and the solid-substrate based immunoassay technology. The low detection limits of goat IgG (gIgG) as 0.042 mu g/mL in buffers, 0.51 mu g/mL in 20-fold diluted whole blood samples and a wide linear range from 0.75 mu g/mL to 60 mu g/mL in blood samples were achieved. To the best of our knowledge, it is the first one-step in situ solid-substrate based immunoassay of whole blood samples with large linear detection range. This development provides a promising platform for a rapid and sensitive immunoassay of various bio-molecules directly in whole blood without tedious separation, washing steps and aggregation problems.