Stability of VIII:C in plasma: the dependence on protease activity and calcium

Thrombosis Research
G A RockD S Palmer

Abstract

Loss of Factor VIII procoagulant activity (VIII:C) following blood collection is a major problem in providing sufficient amounts for therapeutic use and biochemical analyses. We have examined the effects of inhibition of plasma proteases and maintenance of physiological calcium ion on plasma VIII:C stability. The addition of protease inhibitors such as benzamidine, phenylmethylsulfonyl fluoride (PMSF), aprotinin, or soybean trypsin inhibitor (SBTI) to CPD plasma provided no significant protection against decay of VIII:C activity. Neither the rate of decay in the first 24 hours nor the final VIII:C activity observed after storage for 48-72 hours were significantly altered. On the other hand, addition of DFP or heparin to CPD plasma resulted in a marked improvement in VIII:C stability over 24 hours. This demonstrated that these two inhibitors are effective in preventing VIII:C degradation during storage. In addition to protease inhibition, the importance of maintaining physiological calcium ion was demonstrated by 100% stabilization of VIII:C in heparin plasma. Plasma obtained from CPD plus heparin blood could also be stabilized provided free calcium ion levels were restored to physiological concentrations. The inactivation of VI...Continue Reading

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Citations

Oct 15, 1991·Biochemical and Biophysical Research Communications·P R GanzF Luison
Oct 29, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·P Kaersgaard, K A Barington
Aug 2, 1984·The New England Journal of Medicine·G RockD S Palmer
Aug 11, 2006·Clinical and Laboratory Haematology·H YousefG Rock
Jan 1, 1986·Critical Reviews in Clinical Laboratory Sciences·C T Smit Sibinga
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Jan 1, 1984·Scandinavian Journal of Haematology. Supplementum·P R FosterI H Dickson
Jan 1, 1993·Biotechnology Advances·F ManningR O'Kennedy

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