TY - CHAP
T1 - Purification of exosome-like vesicles from urine
AU - Chen, Christopher Y.
AU - Hogan, Marie C.
AU - Ward, Christopher J.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - Density centrifugation
KW - Exosomes
KW - Polycystin
KW - Purification
KW - Tamm Horsfall protein (THP)
UR - http://www.scopus.com/inward/record.url?scp=84875279077&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84875279077&partnerID=8YFLogxK
U2 - 10.1016/B978-0-12-397945-2.00013-5
DO - 10.1016/B978-0-12-397945-2.00013-5
M3 - Chapter
C2 - 23498743
AN - SCOPUS:84875279077
SN - 9780123979452
T3 - Methods in Enzymology
SP - 225
EP - 241
BT - Cilia, Part A
A2 - Marshall, Wallace F.
ER -