Urinary Exosomes for Protein Biomarker Research

Raj, Delfin Albert Amal; Fiume, Immacolata; Capasso, Giovambattista; Pocsfalvi, Gabriella

doi:10.5772/30094

Cited by 2 publications

(2 citation statements)

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“…An additional problem in the isolation of urine samples is the formation of a protein structure of vesiclecrosslinking fibers, for example, from Tamm-Horsfall protein (uromodulin), which can trap uEVs and lead to a reduction in isolation yield. The addition of dithiothreitol (DTT) has been reported as a compound that reduces disulfide bonds, preventing the formation of uromodulin aggregates [2,30,33,34]. It is proposed to add DTT to a pellet obtained after the first centrifugation of urine, and then to incubate and centrifuge it again.…”

Section: Discussionmentioning

confidence: 99%

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Nanoparticle Tracking Analysis of Urinary Extracellular Vesicle Proteins as a New Challenge in Laboratory Medicine

Sałaga-Zaleska,

Kuchta,

Bzoma

et al. 2023

IJMS

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Urinary extracellular vesicle (uEV) proteins may be used as specific markers of kidney damage in various pathophysiological conditions. The nanoparticle-tracking analysis (NTA) appears to be the most useful method for the analysis of uEVs due to its ability to analyze particles below 300 nm. The NTA method has been used to measure the size and concentration of uEVs and also allows for a deeper analysis of uEVs based on their protein composition using fluorescence measurements. However, despite much interest in the clinical application of uEVs, their analysis using the NTA method is poorly described and requires meticulous sample preparation, experimental adjustment of instrument settings, and above all, an understanding of the limitations of the method. In the present work, we demonstrate the usefulness of an NTA. We also present problems encountered during analysis with possible solutions: the choice of sample dilution, the method of the presentation and comparison of results, photobleaching, and the adjustment of instrument settings for a specific analysis. We show that the NTA method appears to be a promising method for the determination of uEVs. However, it is important to be aware of potential problems that may affect the results.

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Section: Discussionmentioning

confidence: 99%

“…Vesicles are released by cells into all body fluids, such as urine, blood, saliva, and amniotic fluid [1,2]. The vesicles secreted into the urine, called urinary extracellular vesicles (uEVs), come from various parts of the urinary system, particularly the kidneys [3].…”

Section: Introductionmentioning

confidence: 99%