Plasmon-Enhanced Biosensing for Multiplexed Profiling of Extracellular Vesicles.

Plasmon-Enhanced Biosensing for Multiplexed Profiling of Extracellular Vesicles.
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DOI:
10.1002/adbi.202000003
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发表时间:
2020-12
影响因子:
4.1
通讯作者:
Im H
Im H
中科院分区:
生物学3区
文献类型:
--
作者:
Min J;Son T;Hong JS;Cheah PS;Wegemann A;Murlidharan K;Weissleder R;Lee H;Im H

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细胞外囊泡(EV)——细胞分泌的纳米级磷脂囊泡——为非侵入性液体活检的分子诊断提供了新的机会。单一 EV 蛋白分析对于研究 EV 作为循环癌症生物标志物可能非常有价值,但由于与有限数量的表位和抗体标记表面积较小相关的检测信号较弱,因此在技术上具有挑战性。在这里,我们报告了一种新的、简单的方法,可以对 EV 标记物进行多重分析,并提高灵敏度。具体来说,我们实施了等离子体增强荧光检测,利用周期性金纳米孔结构激发的表面等离子体共振来放大荧光信号。我们发现多个通道中的荧光信号被放大一个数量级,并且跨膜和囊泡内标记物都可以在单个 EV 水平上检测到。这种方法可以为了解疾病发展和进展过程中 EV 的亚型、异质性和生产动态提供更多见解。等离激元金纳米孔阵列 (nPLEX-FL) 上的等离激元增强荧光检测能够对细胞外囊泡进行灵敏的多重分析。细胞衍生的 EV 被捕获在金纳米孔阵列上,并用荧光团偶联抗体进行标记。然后荧光信号被下面的金纳米孔激发的表面等离子体共振放大,从而实现灵敏的单 EV 检测。
Extracellular vesicles (EVs)—nanoscale phospholipid vesicles secreted by cells—present new opportunities for molecular diagnosis from non-invasive liquid biopsies. Single EV protein analysis could be extremely valuable in studying EVs as circulating cancer biomarkers, but it is technically challenging due to weak detection signals associated with limited amounts of epitopes and small surface areas for antibody labeling. Here, we report a new, simple method that enables multiplexed analyses of EV markers with improved sensitivities. Specifically, we implement plasmon-enhanced fluorescence detection that amplifies fluorescence signals using surface plasmon resonances excited by periodic gold nanohole structures. We showed fluorescence signals in multiple channels are amplified by one order of magnitude, and both transmembrane and intravesicular markers can be detected at the single EV level. This approach could offer additional insight into understanding subtypes, heterogeneity, and production dynamics of EVs during disease development and progression. Plasmon-enhanced fluorescence detection on plasmonic Au nanohole arrays (nPLEX-FL) enables sensitive, multiplexed analysis of extracellular vesicles. Cell-derived EVs are captured on Au nanohole arrays and labeled by fluorophore-conjugated antibodies. The fluorescence signals are then amplified by surface plasmon resonances excited by the underlying Au nanoholes, which lead to sensitive single EV detection.
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影响因子: --
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影响因子: 6.2
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影响因子: 7
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DOI: 10.1007/s11468-013-9660-5
发表时间: 2014
期刊: Plasmonics (Norwell, Mass.)
影响因子: --
作者:
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