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利用表面等离子体共振直接检测炎症触发的内皮细胞衍生的纳米级细胞外囊泡。

Direct detection of nano-scale extracellular vesicles derived from inflammation-triggered endothelial cells using surface plasmon resonance.

机构信息

Hasselt University, Biomedical Research Institute, Hasselt, Belgium.

Maastricht University, Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht, The Netherlands.

出版信息

Nanomedicine. 2017 Jul;13(5):1663-1671. doi: 10.1016/j.nano.2017.03.010. Epub 2017 Mar 30.

Abstract

A major conceptual breakthrough in cell signaling has been the finding of EV as new biomarker shuttles in body fluids. Now, one of the major challenges in using these nanometer-sized biological entities as diagnostic marker is the development of translational methodologies to profile them. SPR offers a promising label-free and real time platform with a high potential for biomarker detection. Therefore, we aimed to develop a uniform SPR methodology to detect specific surface markers on EV derived from patient with CHD. EVs having an approximate size range between 30 and 100 nm (48.5%) and 100-300 nm (51.5%) were successfully isolated. The biomarker profile of EV was verified using immunogold labeling, ELISA and SPR. Using SPR, we demonstrated an increased binding of EV derived from patients with CHD to anti-ICAM-1 antibodies as compared to EV from healthy donors. Our current findings open up novel opportunities for in-depth and label-free investigation of EV.

摘要

细胞信号转导的一个重大概念突破是发现 EV 作为体液中新的生物标志物穿梭物。现在,将这些纳米级生物实体用作诊断标志物的主要挑战之一是开发用于分析它们的转化方法学。SPR 提供了一种有前途的无标记和实时平台,具有高生物标志物检测潜力。因此,我们旨在开发一种统一的 SPR 方法来检测源自 CHD 患者的 EV 上的特定表面标志物。成功分离了大小在 30 到 100nm(48.5%)和 100-300nm(51.5%)之间的 EV。使用免疫金标记、ELISA 和 SPR 验证了 EV 的生物标志物谱。使用 SPR,我们证明与来自健康供体的 EV 相比,源自 CHD 患者的 EV 与抗 ICAM-1 抗体的结合增加。我们目前的发现为深入、无标记的 EV 研究开辟了新的机会。

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