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Updated: May 30, 2026

Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay
Published on: September 9, 2012
Thrombomodulin-dependent effect of factor VLeiden mutation on factor XIII activation
Zsuzsa Koncz1, Zsuzsa Bagoly, Gizella Haramura
1Clinical Research Center University of Debrecen, Medical and Health Science Center, Debrecen, Hungary.
Introduction:
Factor V Leiden mutation (FV(Leiden)) is associated with impaired down-regulation of activated FV procoagulant activity and loss of FV anticoagulant function that result in an increased risk of venous thromboembolism. As the downstream effects of FV(Leiden) on clot formation and fibrinolyis have only partially been revealed, we investigated its effect on the activation of factor XIII (FXIII) and the cross-linking of fibrin.
Methods:
In the plasma samples of fifteen healthy individuals with known FV genotypes coagulation was initiated by recombinant human tissue factor and phospholipids with or without recombinant human thrombomodulin (rhTM). FV deficient plasma supplemented with purified wild type FV or FV(Leiden) were also investigated. Clots were recovered and analyzed by SDS-PAGE and quantitative densitometric evaluation of Western blots.
Results:
rhTM considerably delayed the activation of FXIII in the plasma from FV wild type individuals. This effect of rhTM was significantly impaired in the plasma from FV(Leiden) carriers. The results were confirmed in experiments with FV deficient plasma supplemented by FV prepared from wild type individuals or FV(Leiden) homozygotes. Fibrin γ-chain dimerization was also considerably delayed by rhTM in plasma samples from individuals without Leiden mutation, but not in plasma samples from FV(Leiden) heterozygotes or homozygotes. The difference between heterozygotes and homozygotes was not statistically significant.
Conclusion:
The highly diminished delaying effect of TM on FXIII activation and on the cross-linking of fibrin in FV(Leiden) carriers might represent a novel mechanism contributing to the increased thrombosis risk of these individuals.
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