Citations 2
Calibration of Flow Cytometers Enables Reproducible Measurements of Extracellular Vesicle Concentrations and Reference Range Establishment
Bettin, Britta , Li, Bo , Falkena, Kim, Leeuwen, Ton G. van , Gollwitzer, Christian , Varga, Zoltán , Antović, Jovan P. , Berckmans, Pascale , Buzás, Edit I. , Cook, Sean, Dignat‐George, Françoise , Lacroix, Romaric , Maaninka, Katariina , Menezes, Erika G. Marques de , Meyer, Annette, Mizenko, Rachel R. , Nelissen, Inge , Norris, Philip J. , Pink, Desmond , Poudel, Sumeet , Robert, Stéphane, Siljander, Pia , Tang, Vera A. , Broeck, Tina Van Den, Bettin, Britta A , van Leeuwen, Ton G , Ajzenberg, Nadine , Berckmans, Pascale , Buzas, Edit I , Carney, Randy P , Dignat-George, Françoise , Faille, Dorothee , Giebel, Bernd , Jones, Jennifer C , Lacroix, Romaric , Lannigan, Joanne , Lucien, Fabrice , Maaninka, Katariina , de Menezes, Erika G Marques , Mizenko, Rachel R , Nelissen, Inge , Pink, Desmond , Poudel, Sumeet , Siljander, Pia R-M , Tang, Vera A , Tertel, Tobias , Wang, Lili , Welsh, Joshua A , Nieuwland, Rienk , van der Pol, Edwin , Cook, Sean M, Giebel, Bernd , Jones, Jennifer , Kim, Yohan , Lucien, Fabrice , Meyer, Annette, Nolan, John, Welsh, Joshua A , Nieuwland, Rienk
Journal of Extracellular Vesicles | December 15, 2025 | Volume: 14 | Issue: 12 | Pages: No page available
ABSTRACT The concentration of cells is a key component of modern blood tests. Given the biomarker potential of extracellular vesicles (EVs) in blood, we aimed to establish reference ranges for blood cell‐derived EVs using flow cytometry. To address the orders‐of‐magnitude variability in reported EV concentrations between different flow cytometers (FCMs), we first validated a calibration methodology to enable reproducible EV concentration measurements. The methodology was evaluated in an interlaboratory comparison study and shows that calibration reduces the median absolute deviation of EV concentrations measured on 25 different FCMs from 67 % to 25 %–31 %. The calibration methodology was then used to determine reference ranges of erythrocyte‐, leukocyte‐, and platelet‐derived EVs in human blood plasma in a cohort of healthy individuals ( n = 224). This study demonstrates that calibration enables comparable concentration measurements of blood cell‐derived EVs, thereby bringing EVs one step closer to clinical applications.
Roadblocks of Urinary EV Biomarkers: Moving Toward the Clinic
Droste, Marvin , Puhka, Maija , Ng, Monica Suet Ying , Álvarez‐Llamas, Gloria , Borràs, Francesc E. , Büscher, Anja , Bussolati, Benedetta , Dear, James W. , Hoorn, Ewout J. , Lenassi, Metka , Pink, Desmond , Tiwari, Swasti , Vizio, Dolores Di , Zarovni, Nataša , Martens‐Uzunova, Elena S. , Erdbrügger, Uta , Ng, Monica S Y , Alvarez-Llamas, Gloria , Borràs, Francesc E , Bussolati, Benedetta , Falcón-Perez, Juan Manual , Lenassi, Metka , Lucien, Fabrice , Tertel, Tobias , Martens-Uzunova, Elena S , Royen, Martin E. van , Blijdorp, Charles J. , Falcón‐Pérez, Juan Manuel , Giebel, Bernd , Grange, Cristina , Leivo, Janne , Llorente, Alicia , Lucien, Fabrice , Mertens, Inge , Mischak, Harald , Yuen, Peter S.T. , Jenster, Guido , Burger, Dylan
Journal of Extracellular Vesicles | July 17, 2025 | Volume: 14 | Issue: 7 | Pages: No page available
ABSTRACT Despite remarkable interest in the biomarker potential of urinary extracellular vesicles (uEVs) and the identification of numerous promising candidates, their clinical translation still presents multiple challenges. The opportunities for successful translation are obvious, yet the main roadblocks on the way have hardly been systematically considered and more coordinated approaches are needed to overcome them. In the present review article, we have identified the most relevant roadblocks of clinical translation of urinary EV‐based biomarkers and discuss possible solutions to overcome them. These roadblocks are categorized as fundamental and technical but also related to development of novel biomarker assays and clinical acceptance. In addition, hurdles within the regulatory approval process are discussed. It is clear that various roadblocks to clinical translation of urinary EV biomarkers exist; however, they are addressable by promoting rigor and reproducibility as well as collaboration between basic and clinical scientists, clinicians, industry and regulatory bodies. Moreover, knowledge of obstacles for assay development and regulatory requirements should already be considered when developing a new biomarker to maximize the chance of successful translation. This review presents not only a status quo, but also a roadmap for the further development of the field.