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

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

Author information

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: matthew.wood@dpag.ox.ac.uk.
12
Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom. Electronic address: Samir.El-Andaloussi@ki.se.

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

PMID:
25659648
DOI:
10.1016/j.nano.2015.01.003
[Indexed for MEDLINE]
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