ORCID
- C. Wynne: 0000-0002-0481-8366
- S. Meaney: 0000-0002-7789-516X
Abstract
Background and Objectives: Platelet-derived extracellular vesicles (PEVs) are submicron, membrane-bound particles released upon platelet activation, with a recognized role in haemostasis, inflammation and immunoregulation. PEVs remain insufficiently characterized in blood products. This study compared four isolation methods to evaluate their impact on PEV yield, purity and characteristics, aiming to identify a practical approach for transfusion service workflows. Materials and Methods: PEVs were isolated from expired single-donor aphaeresis platelet concentrates (n = 12) using methods based on different isolation principles: ultracentrifugation (UC), size exclusion chromatography (SEC), mixed size/charge separation (hybrid) and an affinity-based spin column method (affinity). Size, number and biochemical marker expression of all extracellular vesicle (EV) isolates were assessed. Results: PEVs were successfully isolated by all methods, although at varying yields. The overall size distribution of all methods was similar, although SEC and affinity methods isolated PEVs with the largest diameters. PEV isolated by the affinity method had the lowest lipid:protein ratio, consistent with high purity. No differences in expression of EV marker CD9 or platelet activation marker CD42b were found. Conclusion: Comparison of the physical and biochemical characteristics of the PEVs isolated by each method reveals that the affinity method was superior to other methods. In addition, its simplicity, cost effectiveness and accessibility make it a practical option for blood transfusion services to further explore the role of PEVs.
Keywords
affinity-based separation, blood component production, blood component storage, platelet concentrates, platelet-derived extracellular vesicles, size exclusion chromatography, transfusion medicine, ultracentrifugation
DOI Link
Publication Date
2026-01-01
Publication Title
Vox Sanguinis
Volume
121
Issue
3
ISSN
0042-9007
First Page
245
Last Page
253
Acceptance Date
2026-01-01
Deposit Date
2026-02-16
Funding
The authors gratefully acknowledge the Trinity Translational Medicine Institute for performing nanoparticle tracking analysis, the Centre of Applied Science for Health, Technological University Dublin, Tallaght Campus for providing access to the flow cytometer and the staff at the Irish Blood Transfusion Service for their assistance with the acquisition of platelet concentrates. Y.C. performed the research, collected and analysed the data and wrote the first draft of the manuscript. A.W. designed the research study, supervised the research and reviewed and edited the manuscript. A.L. contributed to production and provision of essential materials and reviewed the manuscript. F.M.G. contributed to project supervision and reviewed and edited the manuscript. C.W. designed the research study, analysed the data, supervised the research and reviewed and edited the manuscript. S.M. designed the research study, analysed the data, supervised the research and reviewed and edited the manuscript.
Creative Commons License
This work is licensed under a Creative Commons Attribution-No Derivative Works 4.0 International License.
Additional Links
Recommended Citation
Chintala, Y.; Waters, A.; McGrath, F.; Lorusso, A.; Wynne, C.; and Meaney, S., "From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood Centre" (2026). Research Outputs: 2025-Present. 27.
https://arrow.tudublin.ie/schbiosro/27