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

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.

SUBMITTER: Kim J 

PROVIDER: S-EPMC4475045 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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

Kim Jongmin J   Shin Hyunwoo H   Kim Jiyoon J   Kim Junho J   Park Jaesung J  

PloS one 20150619 6

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 applicab  ...[more]

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