Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

doi:10.3390/pharmaceutics12090872

Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

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Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

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punktacja MEiN [2020]: 100

title:	Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation
author:	Drożdż Anna , Kamińska Agnieszka , Surman Magdalena , Gonet-Surówka Agnieszka , Jach Robert , Huras Hubert , Przybyło Małgorzata , Stępień Ewa
journal title:	Pharmaceutics
volume:	12
issue:	9
date of publication :	2020
article ID:	872
eISSN:	1999-4923
DOI:	10.3390/pharmaceutics12090872
language:	English
journal language:	English
abstract in English:	Recent years have brought great focus on the development of drug delivery systems based on extracellular vesicles (EVs). Considering the possible applications of EVs as drug carriers, the isolation process is a crucial step. To solve the problems involved in EV isolation, we developed and validated a new EV isolation method—low-vacuum filtration (LVF)—and compared it with two commonly applied procedures—differential centrifugation (DC) and ultracentrifugation (UC). EVs isolated from endothelial cell culture media were characterized by (a) Transmission Electron Microscopy (TEM), (b) Nanoparticle Tracking Analysis (NTA), (c) Western blot and (d) Attenuated Total Reflection Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Additionally, the membrane surface was imaged with Environmental Scanning Electron Microscopy (ESEM). We found that LVF was a reproducible and efficient method for EV isolation from conditioned media. Additionally, we observed a correlation between ATR-FTIR spectra quality and EV and protein concentration. ESEM imaging confirmed that the actual pore diameter was close to the values calculated theoretically. LVF is an easy, fast and inexpensive EV isolation method that allows for the isolation of both ectosomes and exosomes from high-volume sources with good repeatability. We believe that it could be an efficient alternative to commonly applied methods.
keywords in English:	dialysis membrane, ectosomes, exosomes, FTIR, infrared spectroscopy, purification
affiliation:	Wydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiego, Wydział Biologii : Instytut Zoologii i Badań Biomedycznych, Wydział Lekarski : Klinika Położnictwa i Perinatologii, Wydział Lekarski : Klinika Endokrynologii Ginekologicznej, Wydział Chemii : Zakład Chemii Ogólnej
type:	journal article
subtype:	academic paper