Year: 2015
Type: article
Abstract: Recent studies have shown that urinary micro-vesicles (MVs) constitute a novel cellular source of biomarkers for kidney diseases.Although the available ultracentrifugation and ultrafiltration protocols effectively isolate urinary MVs, these procedures are ineffective for MVs isolation from urine containing abundant soluble proteins as found in different kidney diseases.Hence, we present a simple and convenient gel sieving chromatography based method that improves the isolation of MVs from urine containing abundant soluble proteins.Spot urine was collected from type-1 diabetes mellitus patients (albumin to creatinine ratio, ACR>300mg/g, proteins=4+) and nonproteinuria healthy subjects (ACR<30mg/g, proteins=nil).MVs were concentrated using ultracentrifugation followed by gel exclusion column chromatography.Urinary MVs fractions were collected and checked on Western blot using exosome-specific antibodies.The results showed significant removal of soluble proteins from MVs of proteinuria.Hence, this technique represents an easy and cost effective approach that could be efficiently utilized for partial purification of MVs from urine containing high proteins.
Source: MOJ Cell Science & Report
Author: Anuradha K
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W2187053680