title: Purification of exosome-like vesicles from urine.
abstract: Urinary exosome-like vesicles (ELVs), 20-200nm membrane-bound particles shed by renal epithelium, have been shown to interact with the primary cilia of distant epithelial cells of the nephron. These ELVs are emerging as an important source of protein, mRNA, and miRNA biomarkers to monitor renal disease. However, purification of ELVs is compromised by the presence of large amounts of the urinary protein Tamm-Horsfall Protein (THP). THP molecules oligomerize into long, double-helical strands several microns long. These linear assemblies form a three-dimensional gel which traps and sequesters ELVs in any centrifugation-based protocol. Here, we present a purification protocol that separates ELVs from THP and divides urinary ELVs into three distinct populations.
authors.affiliation: Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, USA.
authors.forename: Christopher Y
authors.initials: CY
authors.lastname: Chen
authors.forename: Marie C
authors.initials: MC
authors.lastname: Hogan
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Ward
mesh_terms: D015415:Biomarkers / Q000737:chemistry; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D055354:Exosomes / Q000737:chemistry*; D006801:Humans; D011480:Protease Inhibitors / Q000737:chemistry; D013395:Sucrose; D014551:Urinary Tract / Q000737:chemistry / Q000378:metabolism; D014556:Urine / Q000737:chemistry*; D058949:Uromodulin / Q000302:isolation & purification*; D019459:Urothelium / Q000737:chemistry / Q000378:metabolism
chemical_list: D015415:Biomarkers; D011480:Protease Inhibitors; C548612:UMOD protein, human; D058949:Uromodulin; D013395:Sucrose