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

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

PloS one (2015-06-20)
Jongmin Kim, Hyunwoo Shin, Jiyoon Kim, Junho Kim, Jaesung Park
ABSTRACT

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.

MATERIALS
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