title: Extracellular vesicles are rapidly purified from human plasma by PRotein Organic Solvent PRecipitation (PROSPR).
abstract: Extracellular vesicles (EVs) such as exosomes and microvesicles mediate intercellular communication and regulate a diverse range of crucial biological processes. Host cells that are damaged, infected or transformed release biomarker-containing EVs into the peripheral circulation, where they can be readily accessed for use in diagnostic or prognostic testing. However, current methods of EV isolation from blood plasma are complex and often require relatively large sample volumes, hence are inefficient for widespread use in clinical settings. Here, we report a novel and inexpensive method of rapidly isolating EVs from small volumes of human blood plasma by PRotein Organic Solvent PRecipitation (PROSPR). PROSPR encompasses a rapid three-step protocol to remove soluble proteins from plasma via precipitation in cold acetone, leaving the lipid-encapsulated EVs behind in suspension. This generates higher purity EVs that can then be obtained from filtration or classical ultracentrifugation methods. We foresee that PROSPR-based purification of EVs will significantly accelerate the discovery of new disease biomarkers and the characterization of EVs with potential for clinical applications.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore, 637551.
authors.forename: Xavier
authors.initials: X
authors.lastname: Gallart-Palau
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore, 637551.
authors.forename: Aida
authors.initials: A
authors.lastname: Serra
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore, 637551.
authors.forename: Andrew See Weng
authors.initials: AS
authors.lastname: Wong
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore, 637551.
authors.forename: Sara
authors.initials: S
authors.lastname: Sandin
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|Memory, Aging and Cognition Centre, National University Health System, Singapore.
authors.forename: Mitchell K P
authors.initials: MK
authors.lastname: Lai
authors.affiliation: Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|Memory, Aging and Cognition Centre, National University Health System, Singapore.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Chen
authors.affiliation: Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre Singapore, 11 Hospital Drive, Singapore 169610.
authors.forename: Oi Lian
authors.initials: OL
authors.lastname: Kon
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore, 637551.
authors.forename: Siu Kwan
authors.initials: SK
authors.lastname: Sze
mesh_terms: D015415:Biomarkers; D055252:Cell-Derived Microparticles; D011232:Chemical Precipitation; D055354:Exosomes / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D040901:Proteomics / Q000379:methods; D014461:Ultracentrifugation
chemical_list: D015415:Biomarkers