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Author Spotlight: Advancing EVtrap for High-Throughput Proteomics in Disease Biomarker Discovery

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Simple urine storage protocol for extracellular vesicle proteomics compatible with at-home self-sampling.

L A Erozenci^1,2, T V Pham², S R Piersma²

¹Department of Urology, Cancer Center Amsterdam, Cancer Center Amsterdam, Amsterdam UMC, de Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands.

Scientific Reports

|October 22, 2021

Summary

Urine samples can be stored for up to 8 days at 4°C or room temperature with EDTA without impacting urinary extracellular vesicle (EV) concentration or proteome, enabling home-based biomarker studies.

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A Modified Precipitation Method to Isolate Urinary Exosomes

A Modified Precipitation Method to Isolate Urinary Exosomes

Published on: January 16, 2015

Area of Science:

Biochemistry
Biomarker Discovery
Urology

Background:

Urinary extracellular vesicles (EVs) are promising biomarkers.
Clinical use requires reliable short-term storage protocols, especially for at-home collections.
The impact of delayed processing on urinary EV concentration and proteome is not well understood.

Purpose of the Study:

To evaluate the stability of urinary EVs under different short-term storage conditions.
To assess the effect of storage duration and EDTA preservative on EV concentration and proteome.
To determine the suitability of urine storage for at-home collection and subsequent biomarker analysis.

Main Methods:

Urine samples were stored at 4°C and at room temperature with EDTA.
Storage durations included 0, 2, 4, and 8 days.
EVs were isolated using the ME-kit (VN96-peptide).
EV concentration was measured using EVQuant.
Proteome analysis was performed using data-independent acquisition mass spectrometry.

Main Results:

Urinary EV concentration and proteome remained highly stable under both storage conditions.
No significant changes in protein number or quantitative proteomic profiles were observed.
EDTA did not affect urinary EV concentration or the global proteome.
Storage for up to 8 days showed no significant decay in EV concentration or notable effects on the EV-proteome.

Conclusions:

Urine can be stored at 4°C or with EDTA at room temperature for up to 8 days for EV analysis.
These storage protocols support the use of self-collected urine samples for biomarker studies.
Findings facilitate the clinical implementation of urinary EV biomarkers, particularly in home-based settings.