Factor viii purification process

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...

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530413, 530415, 530419, 530420, A61K 3514, C07K 1300

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active

052888533

ABSTRACT:
There is provided in accordance with the practice of this invention a process for separating Factor VIII complex from an impure protein fraction containing Factor VIII complex. An aqueous solution of the impure protein fraction containing Factor VIII complex is applied to a heparin-coupled chromatographic medium, to bind the Factor VIII complex to the medium. The Factor VIII is then recovered from the heparin-coupled chromatographic medium by elution with an aqueous solution comprising CaCl.sub.2 and histidine. The recovered Factor VIII is further purified by precipitation with a solution comprising glycine and NaCl, and washing the resultant precipitate with a solution comprising histidine, glycine, and NaCl to provide a Factor VIII complex solution with a specific activity of about 70 to about 150 units/mg.

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Saundry et al., "Chromatography of vWF on Dextran Sulphate Sepharose", Thrombosis Research, 48, 641-652 (1987).
Harrison et al., "Chromatography of the VIII/vWF Complex on Dextran Sulphate Sepharose", Thrombosis Research, 50, 295-304 (1988).
Roberts et al., "von Willebrand Factor Binds Specifically to Sulfated Glycolipids", J. Biol. Chem., 261, 3306-3309 (1986).
Hamer et al., "Human factor VIII: purification from commercial factor VIII concentrate, characterization, identification and radiolabeling", Biochim. et Biophys. Acta, 873, 356-366 (1986).
Madaras et al., "Isolation and Insolubilisation of Human F VIII by Affinity Chromatography", Haemostasis, 7, 321-331 (1978).
Flow Charts I and II, which relate to processes previously used for producing Factor VIII.

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