Journal
ANALYTICAL CHEMISTRY
Volume 86, Issue 15, Pages 7229-7233Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac501801m
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Funding
- Swiss National Science Foundation
- European Community [FP7-KBBE-2013-613879]
- Gebert Ruf Stiftung [GRS-038/13]
- Marie Curie Actions
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Cells secrete extracellular vesicles (EVs) into their microenvironment that act as mediators of intercellulard communication under physiological conditions and in this context also actively participate in spreading various diseases. Large efforts are currently made to produce reliable EV samples and to develop, improve, and standardize techniques allowing their biophysical characterization. Here, we used ultrafiltration and size-exclusion chromatography for the isolation and a model-free fluorescence fluctuation analysis for the investigation of the physical and biological properties of EVs secreted by mammalian cells. Our purification strategy produced enriched samples of morphologically intact EVs free of extravesicular proteins and allowed labeling of marker molecules on the vesicle surface for single-vesicle analysis with single-molecule sensitivity. This novel approach provides information on the distribution profile of both EV size and relative expression level of a specific exosomal marker, deciphering the overall heterogeneity of EV preparations.
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