Assessing the individual roles of FII, FV, and FX activity in the thrombin generation process

Cuicui Bai, Joke Konings, Marisa Ninivaggi, Marcus Lancé, Bas de Laat, Romy de Laat-Kremers

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


Thrombin generation (TG) is known as a physiological approach to assess the hemostatic function. Although it correlates well with thrombosis and bleeding, in the current setup it is not sensitive to the effects of fluctuations in single coagulation factors. We optimized the calibrated automated thrombinography (CAT) method to quantify FII, FV and FX activity within the coagulation system. The CAT assay was fine-tuned for the assessment of FII, FV and FX by diluting the samples in FII-, FV-, or FX-deficient plasma, respectively, and measuring TG. Plasma FII levels correlated linearly with the ETP up to a plasma concentration of 100% FII. FV and FX levels correlated linearly with the peak height up to a plasma level of 2.5% FV and 10% FX, respectively. Sensitized CAT protocols were designed by adding a fixed volume of a pre-diluted patient sample to FII, FV, and FX deficient plasma in TG experiments. This approach makes the TG measurement dependent on the activity of the respective coagulation factor. The ETP or peak height were quantified as readouts for the coagulation factor activity. The intra- and inter-assay variation coefficients varied from 5.0 to 8.6%, and from 3.5 to 5.9%, respectively. Reference values were determined in 120 healthy subjects and the assays were clinically validated in 60 patients undergoing coronary artery bypass grafting (CABG). The sensitized CAT assays revealed that the contribution of FII, FV, and FX to the TG process was reduced after CABG surgery, leading to reduced prothrombin conversion and subsequently, lower TG.

Original languageEnglish
Article number1000812
JournalFrontiers in Cardiovascular Medicine
Publication statusPublished - 20 Sept 2022


  • calibrated automated thrombinography
  • coagulation factors
  • factor V
  • factor X
  • prothrombin
  • thrombin generation


Dive into the research topics of 'Assessing the individual roles of FII, FV, and FX activity in the thrombin generation process'. Together they form a unique fingerprint.

Cite this