Comparison of assays measuring extracellular vesicle tissue factor in plasma samples: communication from the ISTH SSC Subcommittee on Vascular Biology
Permanent link
https://hdl.handle.net/10037/36574Date
2024-06-24Type
Journal articleTidsskriftartikkel
Peer reviewed
Author
Bonifay, Amandine; Mackman, Nigel; Hisada, Yohei; Sachetto, Ana Teresa Azevedo; Hau, Chi; Gray, Elaine; Hogwood, John; Aharon, Anat; Badimon, Lina; Barile, Lucio; Baudar, Justine; Beckmann, Lennart; Benedikter, Birke; Bolis, Sara; Bouriche, Tarik; Brambilla, Marta; Burrello, Jacopo; Camera, Marina; Campello, Elena; Ettelaie, Camille; Faille, Dorothée; Featherby, Sophie; Franco, Corentin; Guldenpfennig, Maite; Hansen, John Bjarne; Judicone, Coralie; Kim, Yohan; Kristensen, Soren Risom; Laakmann, Katrin; Langer, Florian; Latysheva, Nadezhda; Lucien, Fabrice; de Menezes, Erika Marques; Mullier, François; Norris, Philip; Nybo, Jette; Orbe, Josune; Østerud, Bjarne; Paramo, Jose A.; Radu, Claudia M.; Roncal, Carmen; Samadi, Nazanin; Snir, Omri; Suades, Rosa; Ullsten-Wahlund, Casper Jan Elis; Chareyre, Corinne; Abdili, Evelyne; Martinod, Kimberly; Thaler, Johannes; Dignat-George, Françoise; Nieuwland, Rienk; Lacroix, RomaricAbstract
Objectives - The International Society on Thrombosis and Haemostasis Scientific and Standardization Committee Subcommittee on Vascular Biology initiated a multicenter study to compare the sensitivity, specificity, and reproducibility of these assays.
Methods - Platelet-depleted plasma samples were prepared from blood of healthy donors. The plasma samples were spiked either with EVs from human milk or EVs from TF-positive and TF-negative cell lines. Plasma was also prepared from whole human blood with or without lipopolysaccharide stimulation. Twenty-one laboratories measured EV-TF activity and antigen in the prepared samples using their own assays representing 18 functional and 9 antigenic assays.
Results - There was a large variability in the absolute values for the different EV-TF activity and antigen assays. Activity assays had higher specificity and sensitivity compared with antigen assays. In addition, there was a large intra-assay and interassay variability. Functional assays that used a blocking anti-TF antibody or immunocapture were the most specific and sensitive. Activity assays that used immunocapture had a lower coefficient of variation compared with assays that isolated EVs by high-speed centrifugation.
Conclusion - Based on this multicenter study, we recommend measuring EV-TF using a functional assay in the presence of an anti-TF antibody.