Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties
Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties
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DOI：
10.1016/j.nano.2015.01.003
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发表时间：
2015-05-01
影响因子：
5.4
通讯作者：
Andaloussi, Samir E. L.
中科院分区：
文献类型：
--
作者：
Nordin, Joel Z.;Lee, Yi;Andaloussi, Samir E. L.
细胞外囊泡（EV）是介导RNA和蛋白质的细胞间转移的天然纳米颗粒，并且具有极大的医学兴趣;用作新型生物标志物和潜在的治疗剂。然而，对于从各种生物液体中分离高产和高纯度EV的最合适方法几乎没有共识。在这里，我们描述了一个系统的EV纯化的两个协议之间的比较：超滤，随后的液相色谱（UF-LC）和微分超离心（UC）。一个显着更高的EV产量导致UF-LC相比，UC，而不影响囊泡蛋白质的组成。重要的是，我们提供了新的证据表明，与UC纯化的EV相比，UF-LC纯化的EV的生物物理特性得到保留，导致不同的体内生物分布，在肺中的累积较少。最后，我们表明，UF-LC是可扩展的，适用于从复杂的培养基类型，如干细胞培养基中分离EV，这对未来涉及EV的临床应用具有巨大意义。(C)2015作者爱思唯尔公司出版
Extracellular vesicles (EVs) are natural nanoparticles that mediate intercellular transfer of RNA and proteins and are of great medical interest; serving as novel biomarkers and potential therapeutic agents. However, there is little consensus on the most appropriate method to isolate high-yield and high-purity EVs from various biological fluids. Here, we describe a systematic comparison between two protocols for EV purification: ultrafiltration with subsequent liquid chromatography (UF-LC) and differential ultracentrifugation (UC). A significantly higher EV yield resulted from UF-LC as compared to UC, without affecting vesicle protein composition. Importantly, we provide novel evidence that, in contrast to UC-purified EVs, the biophysical properties of UF-LC-purified EVs are preserved, leading toadifferent in vivo biodistribution, with less accumulation in lungs. Finally, we show that UF-LC is scalable and adaptable for EV isolation from complex media types such as stem cell media, which is of huge significance for future clinical applications involving EVs. (C) 2015 The Authors. Published by Elsevier Inc.