Size-Exclusion Chromatography-based isolation minimally alters Extracellular Vesicles' characteristics compared to precipitating agents
Gámez-Valero, Ana (Institut Germans Trias i Pujol. Hospital Universitari Germans Trias i Pujol)
Monguió Tortajada, Marta (Institut Germans Trias i Pujol)
Carreras-Planella, Laura (Institut Germans Trias i Pujol)
Franquesa, Marcella (Institut Germans Trias i Pujol)
Beyer, Katrin (Institut Germans Trias i Pujol)
Borràs i Serres, Francesc Enric (Institut Germans Trias i Pujol)

Date:	2016
Abstract:	Extracellular vesicles (EVs) have become an attractive field among the scientific community. Yet, a major challenge is to define a consensus method for EVs isolation. Ultracentrifugation has been the most widely used methodology but rapid methods, including Size Exclusion Chromatography (SEC) and/or precipitating agents such as Polyethylene glycol (PEG) or PRotein Organic Solvent PRecipitation (PROSPR) have emerged. To evaluate the impact of these different methods on the resulting EV preparations, plasma EVs were isolated using SEC, PEG and PROSPR, and their total protein content, NTA and Cryo-electron microscopy profiles, and EV-markers were compared. Also, their effect on recipient cells was tested. Low protein content and Cryo-EM analysis showed that SEC removed most of the overabundant soluble plasma proteins, which were not removed using PEG and partially by PROSPR. Moreover, only SEC allowed the detection of the EV-markers CD9, CD63 and CD81, LGALS3BP and CD5L, suggesting a putative interference of the precipitating agents in the structure/composition of the EVs. Furthermore, PEG and PROSPR-based EV isolation resulted in reduced cell viability in vitro. These results stress that appropriate EV-isolation method should be considered depending on the forthcoming application of the purified EVs.
Grants:	Instituto de Salud Carlos III FISPI13/00050 Instituto de Salud Carlos III RD16/0009 Agència de Gestió d'Ajuts Universitaris i de Recerca 2014-SGR-804 Agència de Gestió d'Ajuts Universitaris i de Recerca 014FIB00649 Agència de Gestió d'Ajuts Universitaris i de Recerca 014BPB00118 Fundació la Marató de TV3 482/U/2014
Note:	Altres ajuts: GC/CES07/015
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Extracellular vesicles ; EV ; EV isolation ; Size Exclusion Chromatography ; Polyethylene glycol ; PRotein Organic Solvent PRecipitation
Published in:	Scientific reports, Vol. 6 (September 2016) , ISSN 2045-2322

DOI: 10.1038/srep33641
PMID: 27640641

9 p, 1.4 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut d'Investigació en Ciencies de la Salut Germans Trias i Pujol (IGTP)
Articles > Research articles
Articles > Published articles