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ExtraPEG

Title: ExtraPEG: A Polyethylene Glycol-Based Method for Enrichment of Extracellular Vesicles.
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Name(s): Rider, Mark A, author
Hurwitz, Stephanie N, author
Meckes, David G, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2016-04-12
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Initially thought to be a means for cells to eliminate waste, secreted extracellular vesicles, known as exosomes, are now understood to mediate numerous healthy and pathological processes. Though abundant in biological fluids, purifying exosomes has been challenging because their biophysical properties overlap with other secreted cell products. Easy-to-use commercial kits for harvesting exosomes are now widely used, but the relative low-purity and high-cost of the preparations restricts their utility. Here we describe a method for purifying exosomes and other extracellular vesicles by adapting methods for isolating viruses using polyethylene glycol. This technique, called ExtraPEG, enriches exosomes from large volumes of media rapidly and inexpensively using low-speed centrifugation, followed by a single small-volume ultracentrifugation purification step. Total protein and RNA harvested from vesicles is sufficient in quantity and quality for proteomics and sequencing analyses, demonstrating the utility of this method for biomarker discovery and diagnostics. Additionally, confocal microscopy studies suggest that the biological activity of vesicles is not impaired. The ExtraPEG method can be easily adapted to enrich for different vesicle populations, or as an efficient precursor to subsequent purification techniques, providing a means to harvest exosomes from many different biological fluids and for a wide variety of purposes.
Identifier: FSU_pmch_27068479 (IID), 10.1038/srep23978 (DOI), PMC4828635 (PMCID), 27068479 (RID), 27068479 (EID), srep23978 (PII)
Grant Number: R15 CA188941, R15CA188941
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4828635.
Subject(s): Cell Fractionation/methods
Cell Line
Centrifugation/methods
Epithelial Cells/drug effects
Extracellular Vesicles/chemistry
Extracellular Vesicles/drug effects
Humans
Polyethylene Glycols/pharmacology
Proteome/analysis
RNA/analysis
Surface-Active Agents/pharmacology
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_27068479
Owner Institution: FSU
Is Part Of: Scientific reports.
2045-2322
Issue: vol. 6

Choose the citation style.
Rider, M. A., Hurwitz, S. N., & Meckes, D. G. (2016). ExtraPEG: A Polyethylene Glycol-Based Method for Enrichment of Extracellular Vesicles. Scientific Reports. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_27068479