Related Experiment Video
Updated: Mar 9, 2026
07:15
Purification of High Yield Extracellular Vesicle Preparations Away from Virus
Published on: September 12, 2019
12.5K
Charge-based precipitation of extracellular vesicles
Maria Chiara Deregibus1, Federico Figliolini2, Sergio D'Antico3
1Department of Medical Sciences, University of Turin, I-10126 Turin, Italy.
International Journal of Molecular Medicine
|December 28, 2016
Summary
A novel charge-based precipitation method efficiently isolates extracellular vesicles (EVs) from biological fluids. This simple technique, using protamine and PEG, offers a cost-effective alternative to ultracentrifugation for EV purification and biomarker discovery.
Area of Science:
- Cell Biology
- Biochemistry
- Biotechnology
Background:
- Extracellular vesicles (EVs) mediate intercellular communication and are crucial in physiological and pathological processes.
- EVs in biological fluids serve as potential biomarkers due to their cell-specific molecular signatures.
- Current EV purification methods, while advancing, present challenges and limitations.
Purpose of the Study:
- To develop and evaluate a novel, simplified method for extracellular vesicle (EV) purification.
- To compare the efficacy of charge-based precipitation with traditional differential ultracentrifugation for EV isolation.
- To assess the quality, yield, and biological activity of EVs purified by the new method.
Main Methods:
- A charge-based precipitation technique using protamine (P) and polyethylene glycol (PEG) was developed for EV isolation.
- EVs were precipitated from serum, saliva, and cell culture supernatants.
- Purified EVs were analyzed for size, exosomal marker expression (CD63, CD9, CD81), RNA recovery, and biological activity.
Main Results:
- Protamine/PEG (P/PEG) precipitation effectively isolated EVs without ultracentrifugation, yielding results comparable to the gold standard.
- EV recovery using P/PEG precipitation was more efficient than ultracentrifugation, with similar EV size and exosomal marker enrichment.
- Purified EVs retained their biological functions, including inducing keratinocyte wound closure and tubular epithelial cell proliferation.
Conclusions:
- Charge-based precipitation offers a simple, cost-effective, and efficient method for isolating EVs from small biological samples.
- This technique avoids the need for expensive equipment, making EV isolation more accessible.
- The P/PEG method yields high-quality EVs suitable for downstream applications, including biomarker analysis and functional studies.
