Management of Tamm-Horsfall Protein for Reliable Urinary Analytics.

ABSTRACT:

Purpose

Urinary extracellular vesicles (uEVs) are a novel source of biomarkers. However, urinary Tamm-Horsfall Protein (THP; uromodulin) interferes with all vesicle isolation attempts, precipitates with normal urinary proteins, thus, representing an unwanted "contaminant" in urinary assays. Thus, the aim is to develop a simple method to manage THP efficiently.

Experimental design

The uEVs are isolated by hydrostatic filtration dialysis (HFD) and treated with a defined solution of urea to optimize release of uEVs from sample. Presence of uEVs is confirmed by transmission electron microscopy, Western blotting, and proteomic profiling in MS.

Results

Using HFD with urea treatment for uEV isolation reduces sample complexity to a great extent. The novel simplified uEV isola

SUBMITTER: Xu X 

PROVIDER: S-EPMC6900072 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature