Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties

Joel Z Nordin; Yi Lee; Pieter Vader; Imre Mäger; Henrik J Johansson; Wolf Heusermann; Oscar P B Wiklander; Mattias Hällbrink; Yiqi Seow; Jarred J Bultema; Jonathan Gilthorpe; Tim Davies; Paul J Fairchild; Susanne Gabrielsson; Nicole C Meisner-Kober; Janne Lehtiö; C I Edvard Smith; Matthew J A Wood; Samir El Andaloussi

doi:10.1016/j.nano.2015.01.003

Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties

Nanomedicine. 2015 May;11(4):879-83. doi: 10.1016/j.nano.2015.01.003. Epub 2015 Feb 4.

Authors

Joel Z Nordin¹, Yi Lee², Pieter Vader², Imre Mäger³, Henrik J Johansson⁴, Wolf Heusermann⁵, Oscar P B Wiklander¹, Mattias Hällbrink⁶, Yiqi Seow⁷, Jarred J Bultema⁸, Jonathan Gilthorpe⁹, Tim Davies¹⁰, Paul J Fairchild¹⁰, Susanne Gabrielsson⁸, Nicole C Meisner-Kober⁵, Janne Lehtiö⁴, C I Edvard Smith¹, Matthew J A Wood¹¹, Samir El Andaloussi¹²

Affiliations

¹ Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.
² Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom.
³ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom; Institute of Technology, University of Tartu, Tartu, Estonia.
⁴ Cancer Proteomics Mass Spectrometry, Science for Life Laboratory, Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
⁵ Novartis Institutes for Biomedical Research, Basel, Switzerland.
⁶ Department of Neurochemistry, Stockholm University, Stockholm, Sweden.
⁷ Molecular Engineering Laboratory, Proteos, Agency for Science, Technology and Research (A*STAR), Singapore.
⁸ Department of Medicine Solna, Translational Immunology Unit, Karolinska Institutet and University Hospital, Stockholm, Sweden.
⁹ Umeå Centre for Molecular Medicine, Umeå University, Umeå, Sweden.
¹⁰ Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom.
¹¹ Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom. Electronic address: [email protected].
¹² Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom. Electronic address: [email protected].

PMID: 25659648
DOI: 10.1016/j.nano.2015.01.003

Abstract

Extracellular vesicles (EVs) are natural nanoparticles that mediate intercellular transfer of RNA and proteins and are of great medical interest; serving as novel biomarkers and potential therapeutic agents. However, there is little consensus on the most appropriate method to isolate high-yield and high-purity EVs from various biological fluids. Here, we describe a systematic comparison between two protocols for EV purification: ultrafiltration with subsequent liquid chromatography (UF-LC) and differential ultracentrifugation (UC). A significantly higher EV yield resulted from UF-LC as compared to UC, without affecting vesicle protein composition. Importantly, we provide novel evidence that, in contrast to UC-purified EVs, the biophysical properties of UF-LC-purified EVs are preserved, leading to a different in vivo biodistribution, with less accumulation in lungs. Finally, we show that UF-LC is scalable and adaptable for EV isolation from complex media types such as stem cell media, which is of huge significance for future clinical applications involving EVs.

From the clinical editor: Recent evidence suggests extracellular vesicles (EVs) as another route of cellular communication. These EVs may be utilized for future therapeutics. In this article, the authors compared ultrafiltration with size-exclusion liquid chromatography (UF-LC) and ultra-centrifugation (UC) for EV recovery.

Keywords: Biophysical properties; extracellular vesicles; size-exclusion liquid chromatography; ultracentrifugation; ultrafiltration.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell-Derived Microparticles / chemistry*
Cell-Derived Microparticles / ultrastructure*
Chromatography, Gel
HEK293 Cells
Humans
Ultrafiltration

Grants and funding

G-1109/PUK_/Parkinson's UK/United Kingdom