Method of obtaining intermediate purity factor viii

Details Method of obtaining intermediate purity factor viii Method of obtaining intermediate purity factor viii

A - Human Necessities

23

J

195/39.1

A23J 1/06 (2006.01) A61K 35/16 (2006.01)

Patent

CA 1115637

METHOD OF OBTAINING FACTOR VIII ABSTRACT A method of obtaining Factor VIII which comprises adding heparin to freshly obtained blood plasma collected into a calcium chelating anticoagulant or collecting blood plasma by plasma pheresis using heparin and a calcium chelating anticoagulant; freezing the plasma; resolubilizing the plasma; isolating a cryoprecipitate from the plasma; resolubilizing the cryoprecipitate; adding a citrate saline heparin buffer to the resolubilized temperature of from about 0°C for a time in excess of about one hour, whereby Factor VIII present in the cryoprecipitate is insolubilized using heparin precipitable cold insoluble globulin; separating the Factor VIII rich precipitate; and isolating Factor VIII therefrom. The introduction of the cold insoluble globulin (CIg) step to Factor VIII production results in markedly increased yields of Factor VIII in the cryoprecipitate and, as well, in the cold-insoluble globulin obtained from the cryoprecipitate. Using this procedure, 81% of the Factor VIII is recovered in the cryoprecipitate. The cold-insoluble globulin contains 62% of the starting Factor VIII activity. A final recovery of 666 units per litre of starting plasma is therefore obtained and the amount of protein is reduced to less than 1%. In addition, the procedure can be carried out in blood donor centers, although it is also envisaged to be useful in larger scale recovery of Factor VIII procedures.

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