Document Detail

Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator.
MedLine Citation:
PMID:  17229675     Owner:  NLM     Status:  MEDLINE    
Urinary exosomes are excreted from all nephron segments and may serve as biomarkers for classifying renal diseases. Isolation of urinary exosomes by the established ultracentrifugation method has some limitations for use in a clinical laboratory. We sought a rapid and simple way to obtain urinary exosomes. We used a commercially available nanomembrane concentrator to enrich exosomes from urine by centrifugation at 3,000 g for 10-30 min. Urinary exosomal markers tumor susceptibility gene 101, aquaporin-2, neuron-specific enolase, annexin V, angiotensin-converting enzyme, and podocalyxin (PODXL) were recovered from the nanomembrane concentrator and detected by Western blotting, and typical features of urinary vesicles were found by electron microscopy. Exosomal markers were detected in as little as 0.5 ml of urine. By the nanomembrane method, exosomal proteins could be recovered from urine samples frozen at -80 degrees C or refrigerated overnight at 4 degrees C then stored at -80 degrees C. By enriching exosomes we could detect PODXL, a podocyte marker, which decreased by 71% in five male patients with focal segmental glomerulosclerosis and abundant proteinuria. We conclude that 1) use of a nanomembrane concentrator simplifies and accelerates the enrichment of urinary exosomes; and 2) the nanomembrane concentrator can concentrate exosomal proteins from clinical urine samples. This enhanced method may accelerate the translation of urinary exosomal biomarkers from bench to bedside for the diagnosis, classification, and prognostication of renal diseases.
Anita Cheruvanky; Hua Zhou; Trairak Pisitkun; Jeffrey B Kopp; Mark A Knepper; Peter S T Yuen; Robert A Star
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Intramural     Date:  2007-01-16
Journal Detail:
Title:  American journal of physiology. Renal physiology     Volume:  292     ISSN:  1931-857X     ISO Abbreviation:  Am. J. Physiol. Renal Physiol.     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-05-03     Completed Date:  2007-06-18     Revised Date:  2013-06-06    
Medline Journal Info:
Nlm Unique ID:  100901990     Medline TA:  Am J Physiol Renal Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  F1657-61     Citation Subset:  IM    
Renal Diagnostics and Therapeutics Unit, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-1268, USA.
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MeSH Terms
Biological Markers / urine*
Glomerulosclerosis, Focal Segmental / urine*
Membranes, Artificial*
Nanostructures* / standards
Preservation, Biological
Proteinuria / urine*
Sialoglycoproteins / urine
Ultrafiltration / instrumentation*,  standards
Grant Support
Z01 DK043400-08/DK/NIDDK NIH HHS; Z01 HL001285-21/HL/NHLBI NIH HHS; Z99 HL999999/HL/NHLBI NIH HHS
Reg. No./Substance:
0/Biological Markers; 0/Membranes, Artificial; 0/Polymers; 0/Sialoglycoproteins; 0/Sulfones; 0/podocalyxin; 25667-42-9/polyether sulfone

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