Dataset Information

A novel and simple strategy to isolate extracellular vesicles from human plasma and tissue culture medium with high yield and purity

ABSTRACT: Extracellular vesicles (EVs) function in intercellular communication and can be indicative of the biological status of their producing cells, for example in disease. Isolation of EVs from blood or tissue culture medium is instrumental for the discovery of novel EV associated disease biomarkers as well as for functional studies. Current protocols for EV isolation from blood are limited by low recovery yield and in their capacity to remove contaminating particles, including lipoprotein particles and protein aggregates. In this study, we designed and evaluated a three step protocol to isolate EVs from blood or tissue culture media, with high yield and purity. EVs were first collected and separated from the majority of soluble plasma protein by precipitation with polyethylene glycol (PEG). PEG precipitation resulted in near to absolute recovery, while the yield of EVs collected by ultracentrifugation was only 15%. PEG precipitated EVs were then separated from co-precipitated proteins and protein complexes, low density lipoproteins (LDL), and the majority of high density lipoproteins (HDL) by upward displacement into a iohexol density gradients, yielding 493 fold purification relative to total plasma protein with 76% recovery. Finally, remaining HDL and iohexol was effectively removed by size exclusion chromatography (SEC). Near to absolute pure of isolated EVs was confirmed by cryo-electron microscopy.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood

SUBMITTER: Harmjan Vos

LAB HEAD: Willem Stoorvogel

PROVIDER: PXD015283 | Pride | 2021-09-08

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20190724_F1_RM_XGZ_0272.zip	Other
	20190724_F1_RM_XGZ_0272_DG_F1.raw	Raw
	20190724_F1_RM_XGZ_0272_DG_F2.raw	Raw
	20190724_F1_RM_XGZ_0272_DG_F3.raw	Raw
	20190724_F1_RM_XGZ_0272_SEC_F1.raw	Raw

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Publications

A novel three step protocol to isolate extracellular vesicles from plasma or cell culture medium with both high yield and purity.

Zhang Xiaogang X Borg Ellen G F EGF Liaci A Manuel AM Vos Harmjan R HR Stoorvogel Willem W

Journal of extracellular vesicles 20200714 1

Extracellular vesicles (EV) are membrane encapsulated nanoparticles that can function in intercellular communication, and their presence in biofluids can be indicative for (patho)physiological conditions. Studies aiming to resolve functionalities of EV or to discover EV-associated biomarkers for disease in liquid biopsies are hampered by limitations of current protocols to isolate EV from biofluids or cell culture medium. EV isolation is complicated by the >10<sup>5</sup>-fold numerical excess o ...[more]

PMID: 32944179

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