METHOD OF PRODUCING FACTOR VIII, HAVING IMPROVED RATIO FVIII:C/FVIII:AG Russian patent published in 2019 - IPC C07K1/22 C07K14/755 A61K38/37 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to biotechnology, specifically to a method of producing a product of Factor VIII, having an improved ratio of FVIII:C/FVIII:Ag, by chromatography, and to a product which is obtained using said method. Method of producing product Factor VIII, having ratio FVIII:C/FVIII:Ag of at least 0.7 in product Factor VIII, is carried out by chromatography, where at least one chromatographic step is carried out using an affinity chromatographic resin, on which 13 kDa immobilized Fab antibody fragment derived from yeast is directed against Factor VIII molecule. Binding Factor VIII with affinity resin is carried out in conditions of low concentration of salt, equivalent concentration of 0.1–0.5 mol/kg of sodium chloride. Washing of resin is carried out at high concentration of salt in range of 0.3–4 mol/kg of sodium chloride for light chain removal. Elution and collection of Factor VIII is carried out by using concentration of salt in range of 0.5–4 mol/kg of sodium chloride in combination with 40–60 % of ethylene glycol. Produced Factor VIII product contains more than 98 % of monomer and essentially does not have Aggregated Factor VIII, is suitable for treating haemophilia and eliminating formation of Factor VIII product inhibitors.

EFFECT: said product is stable, preserves its biological activity, high content of monomer Factor VIII in the state of freezing and/or lyophilisation for at least 6 months.

15 cl, 23 dwg, 14 tbl, 10 ex