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Proteomics of Extracellular Vesicles Harvested with ExtraPEG

ABSTRACT: Polyethylene glycol (PEG) is commonly used to isolate virus particles from culture medium. This project adapted and optimized the virus procedure for harvesting extracellular vesicles (of similar biophysical properties to viruses) from media. An 8% PEG solution harvested the purest vesicles. Followed by a wash step (ultracentrifugation of concentrated vesicles in saline), this method produced preparations comparable or superior to commonly used methods in terms of purity, abundance, and cost. It is also highly adaptable, allowing the procedure to be used for enriching vesicles from a variety of biological fluids. As proof of principle, extracellular vesicle protein was successfully harvested from HeLa cell medium using the described PEG-based harvest method and analyzed by mass spectrophotometry.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Epithelial Cell, Cell Culture

SUBMITTER: Mark Rider

LAB HEAD: David G. Meckes Jr.

PROVIDER: PXD003209 | Pride | 2016-04-18

REPOSITORIES: Pride

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ExtraPEG: A Polyethylene Glycol-Based Method for Enrichment of Extracellular Vesicles.

Rider Mark A MA Hurwitz Stephanie N SN Meckes David G DG

Scientific reports 20160412

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 ut ...[more]

PMID: 27068479

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