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Peer-reviewedCC BYMolecular oncology

Optimization of seminal extracellular vesicle isolation protocols for cancer biomarker analysis using proteomic profiling.

  • Elen Telumyan1
  • Beatriz Martín-Gracia2,3
  • Eva Bassols-Citores2,3
  • Krizia Sagini2,3
  • Maria K Andersen1
  • Tuula A Nyman4
  • May-Britt Tessem1,5
  • Morten B Rye5,6
  • Alicia Llorente2,3
6 affiliations
  1. 1Department of Circulation and Medical Imaging, NTNU, Trondheim, Norway.
  2. 2Department of Molecular Cell Biology, Institute for Cancer Research, Oslo University Hospital-The Norwegian Radium Hospital, Oslo, Norway.
  3. 3Centre for Cancer Cell Reprogramming, Faculty of Medicine, University of Oslo, Norway.
  4. 4Department of Immunology, University of Oslo and Oslo University Hospital, Norway.
  5. 5Clinic of Surgery, St. Olavs Hospital, Trondheim University Hospital, Norway.
  6. 6Department of Clinical and Molecular Medicine, NTNU, Trondheim, Norway.
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Abstract

Seminal extracellular vesicles (EVs) hold promise as a source of biomarkers for urogenital cancers. In this study, small seminal EVs were isolated from healthy donors by size-exclusion chromatography (SEC) and three ultracentrifugation (UC) protocols, with or without filtration. EV samples prepared by UC had a higher abundance of typical EV-related molecules such as CD9, CD81, and Alix, compared to SEC preparations. However, ribosomal and ER-related proteins were more enriched in SEC samples. Furthermore, the level of several nonvesicular proteins and/or EV-corona proteins was higher in SEC samples. The addition of a filtration step in the UC protocols also reduced the abundance of these proteins. Finally, we found that the isolation protocol affected the presence of prostate cancer-specific proteins in the EV samples, with membrane proteins such as FOLH1, TMPRSS2, and PSCA enriched in samples prepared by UC. In conclusion, our results suggest that SEC and UC enrich for different EV populations, which should be considered when analyzing cancer biomarkers. Moreover, UC samples contained fewer nonvesicular proteins and/or EV-corona proteins, which may enable a more precise detection of EV-based cancer biomarkers.

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Cite

BibTeXRIS
BibTeX
@article{telumyan2026optimization,
  title = {{Optimization of seminal extracellular vesicle isolation protocols for cancer biomarker analysis using proteomic profiling.}},
  author = {Elen Telumyan and Beatriz Martín-Gracia and Eva Bassols-Citores and Krizia Sagini and Maria K Andersen and Tuula A Nyman and May-Britt Tessem and Morten B Rye and Alicia Llorente},
  journal = {Molecular oncology},
  year = {2026},
  month = oct,
  doi = {10.1002/1878-0261.70345},
  url = {https://europepmc.org/article/MED/42910569},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Optimization of seminal extracellular vesicle isolation protocols for cancer biomarker analysis using proteomic profiling.
AU  - Elen Telumyan
AU  - Beatriz Martín-Gracia
AU  - Eva Bassols-Citores
AU  - Krizia Sagini
AU  - Maria K Andersen
AU  - Tuula A Nyman
AU  - May-Britt Tessem
AU  - Morten B Rye
AU  - Alicia Llorente
PY  - 2026
DA  - 2026/10/22
JO  - Molecular oncology
DO  - 10.1002/1878-0261.70345
UR  - https://europepmc.org/article/MED/42910569
AB  - Seminal extracellular vesicles (EVs) hold promise as a source of biomarkers for urogenital cancers. In this study, small seminal EVs were isolated from healthy donors by size-exclusion chromatography (SEC) and three ultracentrifugation (UC) protocols, with or without filtration. EV samples prepared by UC had a higher abundance of typical EV-related molecules such as CD9, CD81, and Alix, compared to SEC preparations. However, ribosomal and ER-related proteins were more enriched in SEC samples. Furthermore, the level of several nonvesicular proteins and/or EV-corona proteins was higher in SEC samples. The addition of a filtration step in the UC protocols also reduced the abundance of these proteins. Finally, we found that the isolation protocol affected the presence of prostate cancer-specific proteins in the EV samples, with membrane proteins such as FOLH1, TMPRSS2, and PSCA enriched in samples prepared by UC. In conclusion, our results suggest that SEC and UC enrich for different EV populations, which should be considered when analyzing cancer biomarkers. Moreover, UC samples contained fewer nonvesicular proteins and/or EV-corona proteins, which may enable a more precise detection of EV-based cancer biomarkers.
SN  - 1574-7891
SN  - 1878-0261
ER  -