ArticlePloS one2023
A sensitive tissue factor activity assay determined by an optimized thrombin generation method.
Article in PloS one, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 5 citations in OpenAlex.
- A Beginner's Guide to Study the Role of Extracellular Vesicles in Coagulation.Journal of extracellular biology · 2026Review
- TFPIα inhibition by andexanet alfa is partially restored by protein S and factor V-short.Blood advances · 2026Article
- Oral contraceptives and tissue factor expression. Comments on "Do combined oral contraceptives induce formation of tissue factor?": reply.Research and practice in thrombosis and haemostasis · 2026Article
- Reporting negative results - the strength of communicating findings.Research and practice in thrombosis and haemostasis · 2026Article
- Do combined oral contraceptives induce formation of tissue factor?Research and practice in thrombosis and haemostasis · 2025Article
- The impact of protein S and tissue factor pathway inhibitor on coagulation, assessed with thrombin generation, in women starting combined oral contraceptives.Research and practice in thrombosis and haemostasis · 2025Article
- Challenges with measuring tissue factor antigen and activity in human plasma.Blood vessels, thrombosis & hemostasis · 2024Review
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundTissue factor (TF) is the principal activator of the coagulation system, but an increased concentration in the blood in cancer and inflammatory diseases has been suggested to play a role increasing the risk of venous thromboembolism. However, measurement of the TF concentration is difficult, and quantitation of activity is the most valid estimation. The objective of this study was to establish a sensitive method to measure TF activity based on thrombin generation.
methodsThe assay is based on thrombin generation (TG) measured on the Calibrated Automated Thrombogram (CAT). Various low concentrations of TF were prepared from reagents containing 1 pM TF and 4 μM phospholipid (PPL), and no TF and 4 μM PPL, and a calibration curve was produced from Lagtime vs TF concentration. TF in blood samples was measured after isolation and resuspension of extracellular vesicles (EVs) in a standard plasma from which EVs had been removed. The same standard plasma was used for the calibrators.
resultsContact activation of the coagulation system was avoided using CTI plasma samples in Monovette tubes. EVs contain procoagulant phospholipids but addition of PPL only reduced lagtime slightly at very low concentrations of TF resulting in overestimation to a lesser extent at 10 fM but no interference at 30 fM or higher. Addition of EVs to the TG analysis induced a small unspecific TF-independent activity (i.e., an activity not inhibited by antibodies against TF) which also may result in a smaller error in estimation of TF activity at very low levels but the effect was negligible at higher concentrations. It was possible to measure TF activity in healthy controls which was found to be 1-6 fM (EVs were concentrated, i.e. solubilized in a lower volume than the original volume plasma). Coefficient of variation (CV) was below 20% at the low level, and below 10% at a level around 100 fM TF. However, the step with isolation of EVs have a higher inherent CV.
conclusionA sensitive and rather precise one-stage TG-based method to measure TF activity has been established.
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