Isolation of High-Purity Extracellular Vesicles by Extracting Proteins Using Aqueous Two-Phase System

>We present a simple and rapid method to isolate extracellular vesicles (EVs) by using a polyethylene glycol/dextran aqueous two-phase system (ATPS). This system isolated more than ~75% of melanoma-derived EVs from a mixture of EVs and serum proteins. To increase the purity of EVs, a batch procedure was combined as additional steps to remove protein contaminants, and removed more than ~95% of the protein contaminants. We also performed RT-PCR and western blotting to verify the diagnostic applicability of the isolated EVs, and detected mRNA derived from melanoma cells and CD81 in isolated EVs.

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Source URL:http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0129760