Isolation of High-Purity Extracellular Vesicles by Extracting Proteins Using Aqueous Two-Phase System |
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| Authors: | Jongmin Kim Hyunwoo Shin Jiyoon Kim Junho Kim Jaesung Park |
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| Affiliation: | 1. Department of Mechanical Engineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.; 2. School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.; University of South Florida College of Medicine, UNITED STATES, |
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| 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. |
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