PXD015283

PXD015283 is an original dataset announced via ProteomeXchange.

Title	A novel and simple strategy to isolate extracellular vesicles from human plasma and tissue culture medium with high yield and purity
Description	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.
HostingRepository	PRIDE
AnnounceDate	2021-09-08
AnnouncementXML	Submission_2021-09-08_13:36:58.173.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Harmjan Vos
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2019-09-02 06:39:48	ID requested
⏵ 1	2021-09-08 13:36:58	announced

Zhang X, Borg EGF, Liaci AM, Vos HR, Stoorvogel W, A novel three step protocol to isolate extracellular vesicles from plasma or cell culture medium with both high yield and purity. J Extracell Vesicles, 9(1):1791450(2020) [pubmed]

submitter keyword: extracellular vesicles

exosomes

isolation

lipoprotein particles

human plasma

Willem Stoorvogel
contact affiliation	Department of biochemistry and cell biology, Faculty of veterinary science, Utrecht University, Utrecht, The Netherlands.
contact email	w.stoorvogel@uu.nl
lab head
Harmjan Vos
contact affiliation	University Medical Center Utrecht Dept. Molecular Cancer Research
contact email	h.r.vos-3@umcutrecht.nl
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/09/PXD015283

PRIDE project URI

• PRIDE
  1. PXD015283
     1. Label: PRIDE project
     2. Name: A novel and simple strategy to isolate extracellular vesicles from human plasma and tissue culture medium with high yield and purity