Evaluation of four commercial ELISAs to measure tissue factor in human plasma

• Published in: Research and practice in thrombosis and haemostasis 2023-03, Vol.7 (3), p.100133, Article 100133
• Main Authors: Sachetto, Ana T.A., Archibald, Sierra J., Bhatia, Ravi, Monroe, Dougald, Hisada, Yohei, Mackman, Nigel
• Format: Article
• Language: English
• Subjects: acute leukemia; coagulation; enzyme-linked immunosorbent assay; extracellular vesicles; Original; plasma; tissue factor
• ISSN: 2475-0379; 2475-0379
• DOI: 10.1016/j.rpth.2023.100133

Under pathological conditions, tissue factor (TF)-positive extracellular vesicles (EVs) are released into the circulation and activate coagulation. Therefore, it is important to identify methods that accurately quantitate levels of TF in plasma. Enzyme-linked immunosorbent assays (ELISAs) are a fast and simple method to quantitate levels of proteins. However, there are several specific challenges with measuring TF antigen in plasma including its low concentration and the complexity of plasma. We aimed to evaluate the ability of 4 commercial ELISAs to measure TF in human plasma. We determined the ability of 4 commercial ELISAs (Imubind, Quantikine, Human SimpleStep, and CD142 Human) to detect recombinant human TF (Innovin) (12.5-100 pg/mL), TF-positive EVs isolated from the culture supernatant from a human pancreatic cancer cell line (57 pg/mL), TF in plasma containing low levels of EV TF activity (1.2-2.6 pg/mL) from lipopolysaccharide-stimulated whole blood, and plasma containing high levels of EV TF activity (151-696 pg/mL) from patients with acute leukemia. The CD142 Human ELISA could not detect recombinant TF. Imubind and Quantikine but not Human SimpleStep detected recombinant TF spiked into plasma and TF-positive EVs isolated from the culture supernatant of a human pancreatic cancer cell line. Quantikine and Imubind could not detect low levels of TF in plasma from lipopolysaccharide-stimulated whole blood. However, Quantikine but not Imubind detected TF in plasma from acute leukemia patients with high levels of EV TF activity. Our results indicate that commercial ELISAs have different abilities to detect TF. Quantikine and Imubind could not detect low levels of TF in plasma, but Quantikine detected TF in plasma with high levels of TF. [Display omitted] •Tissue factor (TF) in human plasma can be measured using activity- or antigen-based assays.•We evaluated the ability of 4 commercial ELISAs to measure TF in human plasma.•Two of the 4 ELISAs detected recombinant TF spiked into human plasma.•One ELISA detected TF in plasma from leukemia patients with high levels of extracellular vesicle TF activity.