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

doi:10.1038/s41598-021-00289-4

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 journal › Article › Academic › peer-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 language	English
Article number	20760
Journal	Scientific Reports
Volume	11
Issue number	1
Early online date	21 Oct 2021
DOIs	https://doi.org/10.1038/s41598-021-00289-4
Publication status	Published - Dec 2021

Bibliographical note

Funding Information:
This work was made possible through the “IMMPROVE” consortium (Innovative Measurements and Markers for Prostate Cancer Diagnosis and Prognosis using Extracellular Vesicles), which is sponsored by an Alpe d’HuZes grant of the Dutch Cancer Society (Grant #EMCR2015-8022). Furthermore, Cancer Center Amsterdam, Netherlands Organization for Scientific Research (NWO Middelgroot, #91116017) are acknowledged for support of the mass spectrometry infrastructure, and the Daniel den Hoed Cancer Treatment Screening Facility (CTSF) for the Opera Phenix HCS.

Publisher Copyright:
© 2021, The Author(s).

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title = "Simple urine storage protocol for extracellular vesicle proteomics compatible with at-home self-sampling",

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.",

author = "Erozenci, {L. A.} and Pham, {T. V.} and Piersma, {S. R.} and Dits, {N. F.J.} and Jenster, {G. W.} and {van Royen}, {M. E.} and Moorselaar, {R. J.A.} and Jimenez, {C. R.} and Bijnsdorp, {I. V.}",

note = "Funding Information: This work was made possible through the “IMMPROVE” consortium (Innovative Measurements and Markers for Prostate Cancer Diagnosis and Prognosis using Extracellular Vesicles), which is sponsored by an Alpe d{\textquoteright}HuZes grant of the Dutch Cancer Society (Grant #EMCR2015-8022). Furthermore, Cancer Center Amsterdam, Netherlands Organization for Scientific Research (NWO Middelgroot, #91116017) are acknowledged for support of the mass spectrometry infrastructure, and the Daniel den Hoed Cancer Treatment Screening Facility (CTSF) for the Opera Phenix HCS. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

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AU - Jenster, G. W.

AU - van Royen, M. E.

AU - Moorselaar, R. J.A.

AU - Jimenez, C. R.

AU - Bijnsdorp, I. V.

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PY - 2021/12

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N2 - 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.

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

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