Activation of Factor IX by Factor XI_a - A Spectrophotometric Assay for Factor IX in Human Plasma^[*]

Summary

The activation of Factor IX by partially purified Factor XI_a was followed by active site titration, gelelectrophoresis and by a spectrophotometric assay. The assay is based on the finding that the rate of Factor X activation in the presence of phospholipid and Ca₂₊ is linear in time and proportional to the amount of Factor IX_a present and can be determined with the chromogenic substrate S2222. Conditions were found that allowed complete activation of Factor IX in human plasma by Factor XI_a. The amount of Factor IX_a present in the plasma sample can be determined with the spectrophotometric assay and is proportional with the amount of plasma present. In plasma from patients receiving vitamin-K antagonists reduced Factor IX activity is found with the spectrophotometric assay, and the new assay method may be useful in monitoring oral anticoagulant therapy.

Keywords

Active site titration of F IX_a - Activation of F IX by F XI_a - Spectrophotometric assay for F IX_a - Chromogenic substrate - Factor XI_a - Factor IX - Hemophilia B

^* Part of this work was presented by G.T. to the Rijksuniversiteit Limburg in partial fulfilment of the requirements for the Ph. D. degree. The chromogenic substrates used in this work were in part donated by KABI AB, Stockholm, Sweden. Part of this work was presented at the VIIIth International Congress on Thrombosis and Haemostasis, Toronto, Canada, July 1981.

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