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Updated: Jul 29, 2026
Protein Digestion, Ultrafiltration, and Size Exclusion Chromatography to Optimize the Isolation of Exosomes from Human Blood Plasma and Serum
Published on: April 13, 2018
Protocol for large-scale, high-yield, high-purity extracellular vesicle purification from human plasma
Saba Zanganeh1, Gabriel F Arias2, Allaura S Cone3
1Joint Department of Biomedical Engineering, The University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Abstract:
Extracellular vesicles (EVs) hold promise as biomarkers and drug delivery vehicles; however, their broader use is limited by the lack of effective and scalable purification methods. We present a protocol for purifying EVs from human plasma without ultracentrifugation. The workflow incorporates tangential flow filtration (TFF) for volume reduction, polyethylene glycol (PEG)-based precipitation for enrichment, nuclease treatment to remove extravesicular nucleic acids, and multimodal and affinity chromatography for contaminant depletion. We further describe procedures for biochemical, biophysical, and functional EV characterization.
