Reproducible and scalable purification of extracellular vesicles using combined bind-elute and size exclusion chromatography.
Reproducible and scalable purification of extracellular vesicles using combined bind-elute and size exclusion chromatography.
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DOI：
10.1038/s41598-017-10646-x
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发表时间：
2017-09-14
影响因子：
4.6
通讯作者：
Andaloussi SE
中科院分区：
文献类型：
--
作者：
Corso G;Mäger I;Lee Y;Görgens A;Bultema J;Giebel B;Wood MJA;Nordin JZ;Andaloussi SE
细胞外囊泡（EV）在细胞间通讯中起着关键作用，并已被证明参与了多种生理和病理过程。EV传统上通过超离心（UC）纯化，然而UC具有局限性，包括导致操作者依赖性产率、EV聚集和改变的EV形态，并且此外是耗时的。在这里，我们表明，与目前的方法相比，市售的结合型尺寸排阻色谱（BE-SEC）柱以高效的方式以高产率（回收率约80%）纯化EV。该技术是可重复的和可扩展的，并且通过基于珠粒的流式细胞术的表面标记物分析揭示了与UC纯化的样品相比高度相似的表达特征。此外，受体细胞中eGFP标记的EV的摄取在BE-SEC和UC样品之间相当。因此，基于BE-SEC的EV纯化方法代表了一项重要的方法学进步，可能有助于EV生物学和治疗应用的稳健且可重复的研究。
Extracellular vesicles (EVs) play a pivotal role in cell-to-cell communication and have been shown to take part in several physiological and pathological processes. EVs have traditionally been purified by ultracentrifugation (UC), however UC has limitations, including resulting in, operator-dependant yields, EV aggregation and altered EV morphology, and moreover is time consuming. Here we show that commercially available bind-elute size exclusion chromatography (BE-SEC) columns purify EVs with high yield (recovery ~ 80%) in a time-efficient manner compared to current methodologies. This technique is reproducible and scalable, and surface marker analysis by bead-based flow cytometry revealed highly similar expression signatures compared with UC-purified samples. Furthermore, uptake of eGFP labelled EVs in recipient cells was comparable between BE-SEC and UC samples. Hence, the BE-SEC based EV purification method represents an important methodological advance likely to facilitate robust and reproducible studies of EV biology and therapeutic application.
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影响因子：
3.7
作者：
Ghosh A;Davey M;Chute IC;Griffiths SG;Lewis S;Chacko S;Barnett D;Crapoulet N;Fournier S;Joy A;Caissie MC;Ferguson AD;Daigle M;Meli MV;Lewis SM;Ouellette RJ
通讯作者：
Ouellette RJ
影响因子：
5.5
作者：
Blom, Hans;Akerblom, Anna;Lundgren, Mats
通讯作者：
Lundgren, Mats
DOI：
10.1083/jcb.101.3.942
发表时间：
1985-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Pan BT;Teng K;Wu C;Adam M;Johnstone RM
通讯作者：
Johnstone RM
影响因子：
5.4
作者：
Admyre, Charlotte;Johansson, Sara M.;Gabrielsson, Susanne
通讯作者：
Gabrielsson, Susanne
影响因子：
20.3
作者：
Barres, Celine;Blanc, Lionel;Vidal, Michel
通讯作者：
Vidal, Michel