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Thromb Haemost 1990; 63(03): 392-402
DOI: 10.1055/s-0038-1645054
Original Articles
Schattauer GmbH Stuttgart

Development of a Heat-Treated Factor VIII/ von Willebrand Factor Concentrate Prepared from Heparinized Plasma

D S Palmer
The Ottawa Centre of the Canadian Red Cross, Blood Transfusion Service, ottawa,Ontario,Canada
,
P R Ganz
+   Departments of Biochemistry and Laboratory Medicine, University of Ottawa, Ottawa, Ontario, Canada
,
H Perkins
The Ottawa Centre of the Canadian Red Cross, Blood Transfusion Service, ottawa,Ontario,Canada
,
D Rosborough
The Ottawa Centre of the Canadian Red Cross, Blood Transfusion Service, ottawa,Ontario,Canada
,
G Rock
*   Department of Medicine, University of Ottawa, Ottawa, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received 11 November 1988

Accepted after revision 07 March 1990

Publication Date:
24 July 2018 (online)

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Summary

A high yield, intermediate purity factor VIII concentrate derived from heparinized plasma has been developed which can be heat-treated at 60 °C, 68 °C or 80 °C/72 h to permit inactivation of viral contaminants which may be present. After cold reprecipitation of the heparinized cryoprecipitate (CRC), the resolubilized CRC precipitate was adjusted to 25-30 mg/ml protein and pH 6.35 ± 0.1 and incubated for 1 h at 8 °C. After centrifugation to remove the precipitated fibrinogen and fibronectin, a factor VIII-rich supernatant can be recovered which contains > 500 units of VIII : C per liter of starting plasma (Method I product) at a purity of 1.5 U/mg protein. Adjusted to 50 mM glycine and pH 6.8, the product can be lyophilized and heat-treated at 60 °C/72 h without a significant loss of VIII :C activity. However, at 68 °C or 80 °C/72 h, temperatures now reported to be more effective in viral inactivation, the recoveries were reduced to 68 and 33% respectively. Significantly improved recoveries after heat-treatment (HT) at 68 °C or 80 °C/72 h were achieved if the 8 °C supernatant product was prepared by a modified procedure (Method II). This further reduces the fibrinogen content of the product while maintaining VIII : C yields > 500 U/l at a purity of 1.9 U/mg. When adjusted to 50 mM glycine and 1-2% (w/v) sucrose (pH 6.8), lyophilized and heat treated at 60 °C, 68 °C or 80 °C/72 h, the VIII :C recoveries of Method II product were 88-100%, 79-84% and 80-83% of pre-HT levels respectively. The yield of VIII : C was > 400 U/l at a purity of 1.6-1.4 U/mg at 1-2% (w/v) sucrose even after the severe heat-treatment at 80 °C. In addition, the von Willebrand factor multimers are similar in size and triplet pattern to those observed in routine cryoprecipitate preparations.

 

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