Purification of exosome-like vesicles from urine

Christopher Y. Chen, Marie C. Hogan, Christopher J. Ward

Research output: Chapter in Book/Report/Conference proceedingChapter

29 Scopus citations

Abstract

Urinary exosome-like vesicles (ELVs), 20-200 nm 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.

Original languageEnglish (US)
Title of host publicationCilia, Part A
EditorsWallace F. Marshall
Pages225-241
Number of pages17
DOIs
StatePublished - Mar 26 2013

Publication series

NameMethods in Enzymology
Volume524
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

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Keywords

  • Density centrifugation
  • Exosomes
  • Polycystin
  • Purification
  • Tamm Horsfall protein (THP)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Chen, C. Y., Hogan, M. C., & Ward, C. J. (2013). Purification of exosome-like vesicles from urine. In W. F. Marshall (Ed.), Cilia, Part A (pp. 225-241). (Methods in Enzymology; Vol. 524). https://doi.org/10.1016/B978-0-12-397945-2.00013-5