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Extracellular vesicles are rapidly purified from human plasma by PRotein Organic Solvent PRecipitation (PROSPR)

机译:通过PRotein有机溶剂沉淀(PROSPR)从人血浆中快速纯化细胞外囊泡

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摘要

Extracellular vesicles (EVs) such as exosomes and microvesicles mediate intercellular communication and regulate a diverse range of crucial biological processes. Host cells that are damaged, infected or transformed release biomarker-containing EVs into the peripheral circulation, where they can be readily accessed for use in diagnostic or prognostic testing. However, current methods of EV isolation from blood plasma are complex and often require relatively large sample volumes, hence are inefficient for widespread use in clinical settings. Here, we report a novel and inexpensive method of rapidly isolating EVs from small volumes of human blood plasma by PRotein Organic Solvent PRecipitation (PROSPR). PROSPR encompasses a rapid three-step protocol to remove soluble proteins from plasma via precipitation in cold acetone, leaving the lipid-encapsulated EVs behind in suspension. This generates higher purity EVs that can then be obtained from filtration or classical ultracentrifugation methods. We foresee that PROSPR-based purification of EVs will significantly accelerate the discovery of new disease biomarkers and the characterization of EVs with potential for clinical applications.
机译:胞外小泡(EVs),如外来体和微泡,介导细胞间的通讯并调节各种关键的生物学过程。受损,感染或转化的宿主细胞会将含有生物标志物的EV释放到外周循环中,在此处可以方便地访问它们以用于诊断或预后测试。但是,当前从血浆中分离出EV的方法很复杂,并且通常需要相对较大的样本量,因此无法在临床环境中广泛使用。在这里,我们报告了一种新的廉价方法,可通过PRotein有机溶剂沉淀(PROSPR)从少量人血浆中快速分离出EV。 PROSPR包含快速三步操作规程,可通过在冷丙酮中沉淀来从血浆中去除可溶性蛋白,而脂质包裹的EV则处于悬浮状态。这会产生更高纯度的电动汽车,然后可以通过过滤或经典的超离心方法获得电动汽车。我们预见,基于PROSPR的电动汽车的纯化将大大加快新疾病生物标记物的发现和电动汽车的表征,具有临床应用潜力。

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