Comparison of assays measuring extracellular vesicle tissue factor in plasma samples: communication from the ISTH SSC Subcommittee on Vascular Biology.
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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, Osterud Bjarne, Paramo Jose A, Radu Claudia M, Roncal Carmen, Samadi Nazanin, Snir Omri, Suades Rosa, Wahlund Casper, Chareyre Corinne, Abdili Evelyne, Martinod Kimberly, Thaler Johannes, Dignat-George Françoise, Nieuwland Rienk, Lacroix Romaric
Аннотация
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.