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Clot waveform analysis (CWA) reveals variability in Factor VIII (FVIII) reagent activity across different assays. The CWA-sTT method offers a more comprehensive evaluation of FVIII hemostatic ability, particularly for concentrates.

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APTT reagentCWAEHL-FVIIIFVIIIsTF/FIX assay

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Area of Science:

  • Biochemistry
  • Hematology
  • Analytical Chemistry

Background:

  • Factor VIII (FVIII) reagent activity exhibits significant variability depending on the assay and activated partial thromboplastin time (APTT) reagents used.
  • Assessing the precise hemostatic function of FVIII reagents is crucial for effective hemophilia treatment.

Purpose of the Study:

  • To investigate the hemostatic ability of various FVIII reagents using clot waveform analysis (CWA).
  • To compare FVIII activity measurements obtained through CWA-APTT, chromogenic assays, and CWA-sTT.

Main Methods:

  • Utilized 12 APTT reagents and 10 FVIII reagents with reference plasma (RP) for CWA.
  • Employed CWA-APTT, a chromogenic assay, and CWA-sTT to measure FVIII activity.
  • Included small amounts of tissue factor-induced FIX activation (sTF/FIXa) and thrombin time (sTT) in the analysis.

Main Results:

  • CWA-APTT showed marked differences in peak time and peak height across FVIII reagents, with generally shortened peak times and lower peak heights compared to RP.
  • FVIII activity measurements varied by method, with CWA-sTT yielding higher values than APTT or chromogenic assays.
  • CWA-sTT detected an elevated second peak in all FVIII reagents, and both CWA-sTF/FIXa and CWA-sTT demonstrated enhancement with APTT reagents.

Conclusions:

  • The hemostatic ability of FVIII reagents is influenced by the choice of APTT reagents.
  • CWA-sTT provides a valuable method for further evaluating the hemostatic capacity of FVIII concentrates, focusing on thrombin burst characteristics.