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Background  Thrombin generation assay ( TGA ) and thrombelastography ( TEG ) are increasingly employed, global, in vitro methods for assessment of the procoagulant potential of plasma/blood and possibly ideally suited tools to monitor, for example, therapy with recombinant factor  VII a ( FVII a). It remains controversial to what extent results obtained with spiked and postinfusion samples reflect the outcome in patients.    Objective  To characterize the  TGA  response to  FVII a in hemophilic plasma and compare with  TEG  data.    Methods  Hemophilia A ( HA ) was induced in platelet‐rich plasma ( PRP ) from healthy volunteers, followed by spiking with  FVII a, γ‐carboxyglutamic acid (Gla)‐domainless  FVII a or V158D/E296V/M298Q‐ FVII a ( FVII a  DVQ  ). Samples were triggered with tissue factor and analyzed by  TGA  and  TEG  in parallel.    Results  Addition of 25 nmol L −1   FVII a to  HA PRP  normalized  TEG  parameters angle and R time, as well as  TGA  lag time, but had poor effects on the thrombin peak height and velocity index. All parameters (at least) returned to normal levels either upon adding a much higher concentration of  FVII a (~1500 nmol L −1 ) or by using the superactive variant  FVII a  DVQ  . Surprisingly, Gla‐domainless derivatives of  FVII a and  FVII a  DVQ   also yielded considerable effects in  HA PRP .    Conclusions  The good general responses to clinically effective concentrations of  FVII a (25 and 75 nmol L −1 ) seen in  TEG  analyses, as well as for  TGA  lag time, were accompanied by far‐from‐normal thrombin peaks. A near‐normal thrombin peak response required the presence of considerably higher  FVII a activity but, intriguingly, relied only marginally on a functional Gla domain (ie, on platelet surface localization).

Limited factor VII a surface localization requirement of the factor VII a–induced overall thrombin generation in platelet‐rich hemophilia A plasma