Abstract A method for recovering Antihemophilic Factor VIII from whole blood or blood plasma or blood plasma fractions freshly collected directly into an anticoagulant selected from the group of heparin, sodium heparin, or mixtures thereof, the heparin being used in the range of 6-8 units/ml of plasma or 3.5-4 units/ml of blood; and either first separating the red cells from the plasma and/or freezing the heparinized plasma, resolubilizing the plasma, isolating a cryoprecipitate from the plasma, resolubilizing the cryoprecipitate, adding a saline heparin solution to the resolubilized cryoprecipitate, incubating the resolubilized cryoprecipitate at a temperature of from about 0° C. to about 4° C. for a time of at least 2 hours, whereby Factor VIII present in the cryoprecipitate is insolubilized using heparin precipitable cold insoluble globulin and the resulting Factor VIII rich precipitate also includes cold insoluble globulin, separating the Factor VIII rich precipitate, and isolating Factor VIII therefrom. This new process permits recovery of a high purity Factor VIII preparation which can easily be made in the blood bank or alternatively, the cold precipitation step can be carried out in a fractionation plant.
In a method of obtaining Factor VIII which comprises (a) 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; (b) freezing the plasma; (c) resolubilizing the plasma; (d) isolating a cryoprecipitate from the plasma; (e) resolubilizing the cryoprecipitate; (f) adding a citrate saline heparin buffer to the resolubilized cryoprecipitate; (g) incubating the buffered, resolubilized cryoprecipitate at a temperature of from about 0° to 10° 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 and the resulting (Factor VIII) rich precipitate also includes cold insoluble globulin; (h) separating the Factor VIII rich precipitate; and (i) isolating Factor VIII therefrom, the improvement consisting of: (1) substituting for step (a) collecting freshly obtained blood plasma or blood plasma fractions directly into an anticoagulant consisting essentially of heparin, sodium heparin, or their mixture, in a proportion of about 6-8 units of heparin per ml of blood plasma, in the absence of a calcium chelating anticoagulant; and (2) substititing for step (f), adding a buffer consisting essentially of a saline heparin solution to the resolubilized cryoprecipitate, in the absence of a citrate buffer.
Claim 2
Dependent Claim
212 words
In a method of obtaining Factor VIII which comprises (a) 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; (b) freezing the plasma; (c) resolubilizing the plasma; (d) isolating a cryoprecipitate from the plasma; (e) resolubilizing the cryoprecipitate; (f) adding a citrate saline heparin buffer to the resolubilized cryoprecipitate; (g) incubating the buffered, resolubilized cryoprecipitate at a temperature of from about 0° to 10° 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 and the resulting (Factor VIII) rich precipitate also includes cold insoluble globulin; (h) separating the Factor VIII rich precipitate; and (i) isolating Factor VIII therefrom, the improvement consisting of: (1) substituting for step (a) collecting freshly obtained blood plasma or blood plasma fractions directly into an anticoagulant consisting essentially of heparin, sodium heparin, or their mixture, in a proportion of about 6-8 units of heparin per ml of blood plasma, in the absence of a calcium chelating anticoagulant; and (2) substititing for step (f), adding a buffer consisting essentially of a saline heparin solution to the resolubilized cryoprecipitate, in the absence of a citrate buffer.
Claim 3
Dependent Claim
38 words
The improvement of claim 1 wherein the blood plasma is obtained from whole blood using plasma pheresis in which the anticoagulant consists essentially of heparin, sodium heparin, or their mixture, in the absence of a calcium chelating anticoagulant.
Claim 4
Dependent Claim
20 words
The improvement of claim 2 or 3 wherein about 8 units of anticoagulant are used per ml of blood plasma.