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

L. A. Erozenci, T. V. Pham, S. R. Piersma, N. F.J. Dits, G. W. Jenster, M. E. van Royen, R. J.A. Moorselaar, C. R. Jimenez*, I. V. Bijnsdorp*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Urinary extracellular vesicles (EVs) have gained increased interest as a biomarker source. Clinical implementation on a daily basis requires protocols that inevitably includes short-term storage of the clinical samples, especially when collected at home. However, little is known about the effect of delayed processing on the urinary EVs concentration and proteome. We evaluated two storage protocols. First, urine stored at 4 °C. Secondly a protocol compatible with at-home collection, in which urine was stored with the preservative EDTA at room temperature (RT). EVs were isolated using the ME-kit (VN96-peptide). For both conditions we explored the effect of storage duration (0, 2, 4 and 8 days) on EV concentration and proteome using EVQuant and data-independent acquisition mass spectrometry, respectively. The urinary EV concentration and proteome was highly stable using both protocols, in terms of protein number and quantitative changes. Furthermore, EDTA does not affect the urinary EV concentration or global proteome. In conclusion, urine can be stored either at 4 °C or with EDTA at RT for up to 8 days without any significant decay in EV concentration or a notable effect on the EV-proteome. These findings open up biomarker studies in urine collected via self-sampling at home.

Original languageEnglish
Article number20760
JournalScientific Reports
Volume11
Issue number1
Early online date21 Oct 2021
DOIs
Publication statusPublished - Dec 2021

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