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US H1,509 HGranted
Heparin enhanced process for separating antihemophilic factor (Factor VIII) and fibronectin from cryoprecipitate
OwnerHarutyun Eran, Duk S. Hwang +1inventors
InventorsHarutyun Eran, Duk S. Hwang, David P. Kosow
Life of this patent
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Abstract
A process is provided for preparing fibronectin and antihemophilic factor (Factor VIII) concentrates from a blood plasma cryoprecipitate.
Claims13
A process for producing a Factor VIII concentrate from blood plasma, the process comprising the steps of:
- (a) obtaining a cryoprecipitate containing Factor VIII from blood plasma;
- (b) dissolving the cryoprecipitate in an aqueous solution containing heparin in an amount sufficient to provide a cryoprecipitate/heparin solution containing from about 30 to about 150 units of heparin per milliliter of solution:
- (c) adding a sufficient amount of a precipitant consisting essentially of PEG to the cryoprecipitate/heparin solution while maintaining the solution at a temperature of from 20.degree. C. to 30.degree. C. to precipitate protein contaminants, leaving a PEG supernatant containing Factor VIII;
- (d) recovering the PEG supernatant;
- and
- (e) recovering Factor VIII from the PEG supernatant.
3 more elements
A process for producing a Factor VIII concentrate from blood plasma, the process comprising the steps of:
- (a) obtaining a cryoprecipitate containing Factor VIII from the blood plasma;
- (b) dissolving the cryoprecipitate in an aqueous solution containing heparin in an amount sufficient to provide a cryoprecipitate/heparin solution containing from about 30 to about 150 units of heparin per milliliter of solution;
- (c) adding a precipitant consisting essentially of PEG to the cryoprecipitate/heparin solution, while maintaining the solution at a temperature of from 20.degree. C. to 30.degree. C., to provide a first solution having a PEG concentration of from about 1% to about 5% and adjusting the pH of the solution to from about 5.9 to about 6.2 to thereby precipitate unwanted protein contaminants from the solution to provide a first PEG supernatant containing Factor VIII;
- (d) recovering the first PEG supernatant;
- (e) adding an additional precipitant consisting essentially of PEG to the first PEG supernatant to bring the PEG concentration to from about 8% to about 16%, while maintaining the solution at a temperature of from about 15.degree. C. to about 30.degree. C., to thereby precipitate Factor VIII from the supernatant;
- (f) recovering and then washing the Factor VIII precipitate in an aqueous solution containing (1) glycine at a concentration of from about 1.0M to about 2.6M, (2) citrate, and (3) heparin at a concentration of from about 10 to about 15 units heparin per milliliter of solution, wherein the wash is maintained at less than about 15.degree. C.;
- and
- (g) recovering the washed Factor VIII precipitate.
5 more elements
A process for producing a Factor VIII concentrate from blood plasma, the process comprising the steps of:
- (a) obtaining a cryoprecipitate containing Factor VIII from blood plasma;
- (b) dissolving the cryoprecipitate in an aqueous solution containing heparin in an amount sufficient to provide a cryoprecipitate/heparin solution containing from about 30 to about 150 units of heparin per milliliter of solution;
- (c) adding a precipitant consisting essentially of PEG to the cryoprecipitate/heparin solution to provide a first solution having a PEG concentration of from about 1% to about 5% and adjusting the pH of the solution to from about 5.9 to about 6.2 to thereby precipitate unwanted protein contaminants from the solution to provide a first PEG supernatant containing Factor VIII;
- (d) recovering the first PEG supernatant;
- (e) adding additional precipitant consisting essentially of PEG to the first PEG supernatant to bring the PEG concentration to from about 8% to about 16% to thereby precipitate Factor VIII from the supernatant;
- (f) recovering and then washing the Factor VIII precipitate in an aqueous solution containing (1) glycine at a concentration of from about 1 to about 2.6M, (2) citrate at a concentration of from about 0.1 to about 0.5M, and (3) heparin at a concentration of from about 10 to about 15 units of heparin per milliliter of solution, wherein the wash is maintained at less than about 15.degree. C.;
- and
- (g) recovering the washed Factor VIII precipitate.
5 more elements
A process for producing a Factor VIII concentrate from blood plasma, the process comprising the steps of:
- (a) obtaining a cryoprecipitate containing Factor VIII from blood plasma;
- (b) dissolving the cryoprecipitate in an aqueous solution containing heparin in an amount sufficient to provide a cryoprecipitate/heparin solution containing about 80 units of heparin per milliliter of solution;
- (c) adding a precipitant consisting essentially of PEG to the cryoprecipitate/heparin solution to provide a first solution having a PEG concentration of about 3% to 4% and adjusting the pH of the solution to about 6.3 to thereby precipitate unwanted protein contaminants from the solution to provide a first PEG supernatant containing Factor VIII;
- (d) recovering the first PEG supernatant;
- (e) adding additional precipitant consisting essentially of PEG to the first PEG supernatant to bring the PEG concentration to about 12% to thereby precipitate Factor VIII from the supernatant;
- (f) recovering and then washing the Factor VIII precipitate in an aqueous solution containing (1) glycine at a concentration of about 1.6M, (2) citrate at a concentration of about 0.2M, and (3) heparin at a concentration of from about 10 to about 15 units of heparin per milliliter of solution, wherein the wash is maintained at less than about 15.degree. C.;
- and
- (g) recovering the washed Factor VIII precipitate.
5 more elements
Dependencies are inferred from the claim text.