Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Low Vacuum Filtration Method as an Alternative Extracellular Vesicle Concentration Method: Comparison with Ultracentrifugation and Differential Centrifugation

Anna Drożdż
ORCID logo ,
Agnieszka Kamińska
ORCID logo ,
Magdalena Surman
ORCID logo ,
Agnieszka Gonet-Surówka
,
Robert Jach
,
Hubert Huras
,
Małgorzata Przybyło
ORCID logo ,
Ewa Łucja Stępień
* ORCID logo
Version 1 : Received: 26 July 2020 / Approved: 27 July 2020 / Online: 27 July 2020 (00:46:38 CEST)

A peer-reviewed article of this Preprint also exists.

Drożdż, A.; Kamińska, A.; Surman, M.; Gonet-Surówka, A.; Jach, R.; Huras, H.; Przybyło, M.; Stępień, E.Ł. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics 2020, 12, 872. Drożdż, A.; Kamińska, A.; Surman, M.; Gonet-Surówka, A.; Jach, R.; Huras, H.; Przybyło, M.; Stępień, E.Ł. Low-Vacuum Filtration as an Alternative Extracellular Vesicle Concentration Method: A Comparison with Ultracentrifugation and Differential Centrifugation. Pharmaceutics 2020, 12, 872.

Abstract

Recent years brought great focus in the field of development of extracellular vesicles (EVs) based drug-delivery systems. Considering possible applications of EVs as a drug carriers the isolation process is a crucial step. To solve problems related with EV isolation, we created and validated a new EVs isolation method – Low Vacuum Filtration (LVF) and compared it with two commonly applied procedures - differential centrifugation (DC) and ultracentrifugation (UC). EVs isolated from endothelial cells culture media have been characterized by a) transmission electron microscopy (TEM) b) nanoparticle tracking analysis (NTA), c) western blot and d) Fourier-Transform Infrared Spectroscopy (FTIR). Additionally, the membrane surface have been imaged with Environmental Scanning Electron Microscopy (ESEM). We showed that LVF is reproducible and efficient method for EVs isolation form conditioned media. Additionally, we observed correlation between ATR-FTIR spectra quality and the EVs and proteins concentration. ESEM imaging confirmed that actual pore diameter are close to the values calculated theoretically. LVF method is an easy, fast and inexpensive EVs isolation method which allows for isolation of both ectosomes and exosomes from high volume sources with good repeatability. We think that it could be an efficient alternative for commonly applied methods.

Keywords

dialysis membrane; ectosomes; exosomes; FTIR; infrared spectroscopy; purification

Subject

Chemistry and Materials Science, Biomaterials

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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