From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood Centre

Y. Chintala; A. Waters; F. McGrath; A. Lorusso; C. Wynne; S. Meaney

doi:10.1111/vox.70172

From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood Centre

Y. Chintala
, A. Waters
, F. McGrath
, A. Lorusso
, C. Wynne
, S. Meaney

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Number of pages	8
Journal	Vox Sanguinis
DOIs	https://doi.org/10.1111/vox.70172
Publication status	Published - 26 Jan 2026

Keywords

affinity-based separation
blood component production
blood component storage, platelet concentrates
ultracentrifugation
platelet-derived extracellular vesicles
size exclusion chromatography
transfusion medicine

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From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood CentreFinal published version, 1.5 MBLicence: CC BY-NC-ND

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title = "From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood Centre",

abstract = "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, ultracentrifugation, platelet-derived extracellular vesicles, size exclusion chromatography, transfusion medicine",

author = "Y. Chintala and A. Waters and F. McGrath and A. Lorusso and C. Wynne and S. Meaney",

year = "2026",

month = jan,

day = "26",

doi = "10.1111/vox.70172",

language = "English",

journal = "Vox Sanguinis",

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T1 - From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood Centre

AU - Chintala, Y.

AU - Waters, A.

AU - McGrath, F.

AU - Lorusso, A.

AU - Wynne, C.

AU - Meaney, S.

PY - 2026/1/26

Y1 - 2026/1/26

N2 - 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.

AB - 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.

KW - affinity-based separation

KW - blood component production

KW - blood component storage, platelet concentrates

KW - ultracentrifugation

KW - platelet-derived extracellular vesicles

KW - size exclusion chromatography

KW - transfusion medicine

U2 - 10.1111/vox.70172

DO - 10.1111/vox.70172

M3 - Article

SN - 0042-9007

JO - Vox Sanguinis

JF - Vox Sanguinis

ER -

From platelets to particles: Optimizing extracellular vesicle isolation in a National Blood Centre

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