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Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay
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Published on: September 9, 2012

The "normal" factor VIII concentration in plasma.

Saulius Butenas1, Behnaz Parhami-Seren, Anetta Undas

  • 1Department of Biochemistry, College of Medicine, University of Vermont, Burlington, VT 05446, USA. sbutenas@uvm.edu

Thrombosis Research
|May 11, 2010
PubMed
Summary
This summary is machine-generated.

Factor VIII antigen (FVIIIag) levels are higher than activity-based measurements in plasma. Measuring FVIIIag provides a more accurate estimation of real FVIII concentrations, accounting for von Willebrand factor influences.

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Area of Science:

  • Hematology
  • Clinical Chemistry
  • Biochemistry

Background:

  • Quantifying factor VIII (FVIII) activity using assays is complex.
  • Assay results are affected by methodology, reagents, and plasma protein concentrations, notably von Willebrand factor (VWF).

Purpose of the Study:

  • To compare FVIII concentrations determined by activity-based assays versus immunoassay.
  • To assess the impact of plasma dilution on FVIII clotting activity (FVIIIc).

Main Methods:

  • Utilized activated partial thromboplastin time (APTT), chromogenic assay (Coatest), and two in-house immunoassays.
  • Employed albumin-free recombinant FVIII as the calibrator across all assays.
  • Investigated plasma dilutions (4-fold, 10-fold, 25-fold) to evaluate FVIIIc in healthy individuals.

Main Results:

  • In healthy individuals, FVIII antigen (FVIIIag) levels (1.22 nM) were significantly higher than FVIII clotting activity (FVIIIc; 0.65 nM) and chromogenic assay (FVIIIch; 0.50 nM) values.
  • Observed a positive correlation between FVIIIag and VWF antigen (VWFag) (R²=0.20).
  • Plasma dilution increased FVIIIc estimates, with 25-fold dilution yielding FVIIIc closest to FVIIIag.

Conclusions:

  • FVIII antigen levels consistently exceed FVIII clotting activity and chromogenic measurements in plasma.
  • Measuring FVIII antigen (FVIIIag) offers a reliable method for estimating true FVIII concentrations in plasma.