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

By J. Nordin, Y. Lee, P. Vader, Imre Mäger, H. Johansson, Wolf Heusermann, O. Wiklander, M. Hällbrink, Y. Seow, J. Bultema and 9 more. Nanomedicine: Nanotechnology, Biology and Medicine, 2015

603 citations recorded by the source corpus.

UNLABELLED 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.

Subfields: Biomedical Engineering, Molecular Biology, Pulmonary and Respiratory Medicine

View Paper: https://www.semanticscholar.org/paper/ae4e6cacabd862d241b10adccbd3eab32dbd0dde

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Biomedical Engineering

1. Hina Kalra
2. Suresh Mathivanan

In Molecular Biology

1. C. Théry
2. Hina Kalra
3. Suresh Mathivanan
4. G. Drummen
5. P. Vader
6. Jan Lötvall
7. Ramesh C. Gupta
8. M. Wood

In Pulmonary and Respiratory Medicine

1. R. Corteling
2. I. Colao
3. I. Wall
4. D. Bracewell

Citations in the corpus

• Focus on Extracellular Vesicles: Introducing the Next Small Big Thing (2016, background): “Finally, specific purification protocols after ultracentrifugation not only affect EV purity and yield, but often lead to protein loss in the preparation [163,164].”
• Extracellular Vesicle Heterogeneity: Subpopulations, Isolation Techniques, and Diverse Functions in Cancer Progression (2018, background): “, vesicles disruption, aggregation, and co-purification of non-EV material (89, 90)].”
• Manufacturing Exosomes: A Promising Therapeutic Platform. (2018, positive): “an improved biodistribution of the EVs when injected into mice: fewer EVs accumulated in the lungs, likely due to the reduction of aggregation and damage to the exosomes during the UF-LC steps compared with ultracentrifugation [69].”
• Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey (2016, positive): “It is possible that different EV isolation methods may preferentially isolate different EV populations, with different size, protein, RNA and functional characteristics (6,13,20).”
• Applying extracellular vesicles based therapeutics in clinical trials – an ISEV position paper (2015, methodology): “Established technologies to determine (average) EV sizes and concentrations are electron microscopy (EM), dynamic light scattering (DLS) (317), nanoparticle tracking analysis (NTA) (313,318), resistive pulse sensing (319) and fluorescence correlation spectroscopy (306).”