重组细胞外囊泡的产生和应用作为生物参考物质。

The generation and use of recombinant extracellular vesicles as biological reference material.

机构信息

Laboratory of Experimental Cancer Research, Department of Human Structure and Repair, Ghent University, Ghent, 9000, Belgium.

Cancer Research Institute Ghent, Ghent, 9000, Belgium.

出版信息

Nat Commun. 2019 Jul 23;10(1):3288. doi: 10.1038/s41467-019-11182-0.

DOI:10.1038/s41467-019-11182-0

PMID:31337761

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6650486/

Abstract

Recent years have seen an increase of extracellular vesicle (EV) research geared towards biological understanding, diagnostics and therapy. However, EV data interpretation remains challenging owing to complexity of biofluids and technical variation introduced during sample preparation and analysis. To understand and mitigate these limitations, we generated trackable recombinant EV (rEV) as a biological reference material. Employing complementary characterization methods, we demonstrate that rEV are stable and bear physical and biochemical traits characteristic of sample EV. Furthermore, rEV can be quantified using fluorescence-, RNA- and protein-based technologies available in routine laboratories. Spiking rEV in biofluids allows recovery efficiencies of commonly implemented EV separation methods to be identified, intra-method and inter-user variability induced by sample handling to be defined, and to normalize and improve sensitivity of EV enumerations. We anticipate that rEV will aid EV-based sample preparation and analysis, data normalization, method development and instrument calibration in various research and biomedical applications.

摘要

近年来，人们越来越关注细胞外囊泡（EV）的研究，旨在深入了解其生物学特性、开发诊断方法和治疗手段。然而，由于生物体液的复杂性以及样本制备和分析过程中引入的技术差异，EV 数据的解读仍然具有挑战性。为了理解和减轻这些限制，我们生成了可追踪的重组 EV（rEV）作为生物参考材料。通过采用互补的表征方法，我们证明 rEV 是稳定的，并且具有与样本 EV 特征相符的物理和生化特性。此外，rEV 可以使用常规实验室中现有的荧光、RNA 和蛋白质技术进行定量。向生物体液中添加 rEV 可以确定常用 EV 分离方法的回收率效率，定义由样本处理引起的方法内和用户间的变异性，并对 EV 计数进行标准化和提高其灵敏度。我们预计 rEV 将有助于基于 EV 的样本制备和分析、数据标准化、方法开发和仪器校准，适用于各种研究和生物医学应用。