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Related Experiment Videos

A method for measuring activated factor VIII in plasma.

H Kessels1, S Béguin, R Wagenvoord

  • 1Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.

Thrombosis and Haemostasis
|October 1, 1991
PubMed
Summary

A new method quantifies activated factor VIII (VIIIa) in plasma by measuring factor X activation. This technique provides a sensitive and reliable way to assess factor VIIIa concentrations.

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Area of Science:

  • Biochemistry
  • Hematology

Background:

  • Accurate measurement of activated factor VIII (VIIIa) is crucial for diagnosing and managing coagulation disorders.
  • Existing methods may be affected by interfering factors in plasma.

Purpose of the Study:

  • To develop a quantitative method for measuring activated factor VIII (VIIIa) concentration in plasma.
  • To establish a reliable and specific assay for factor VIIIa.

Main Methods:

  • Utilizing the cofactor activity of factor VIIIa in accelerating factor X activation by factor IXa, phospholipid, and calcium.
  • Measuring the rate of factor X activation over time using a chromogenic substrate.
  • Employing a high concentration of chromogenic substrate to simultaneously inhibit factor Xa activity and measure substrate conversion.

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Main Results:

  • Factor X activation velocity was found to be linearly proportional to factor VIIIa concentration under specific conditions.
  • Optical density increased as a parabolic function of time, allowing factor VIIIa concentration to be derived from the quadratic coefficient.
  • The method demonstrated specificity for factor VIIIa, with no influence from extrinsic factor X activators.

Conclusions:

  • A sensitive and reliable method for assessing factor VIIIa concentrations in plasma has been developed.
  • The assay relies on the simultaneous inhibition and measurement of factor Xa via a high concentration of chromogenic substrate.
  • This technique offers a valuable tool for clinical diagnostics and research in hemostasis.