title: 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.
authors.affiliation: Department of Mechanical Engineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.
authors.forename: Jongmin
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Mechanical Engineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.
authors.forename: Hyunwoo
authors.initials: H
authors.lastname: Shin
authors.affiliation: School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.
authors.forename: Jiyoon
authors.initials: J
authors.lastname: Kim
authors.affiliation: School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.
authors.forename: Junho
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Mechanical Engineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea; School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Gyeong-buk, Republic of Korea.
authors.forename: Jaesung
authors.initials: J
authors.lastname: Park
mesh_terms: D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D000067128:Extracellular Vesicles; D059625:Liquid-Liquid Extraction / Q000379:methods; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry; D011506:Proteins / Q000302:isolation & purification*; D014461:Ultracentrifugation
chemical_list: D011506:Proteins; D011092:Polyethylene Glycols