Preparation method of high-purity human coagulation factor IX

Abstract

The invention relates to a preparation method of a high-purity human coagulation factor IX, which comprises the following steps: melting refrigerated plasma, and carrying out low-temperature centrifugation; adsorbing with a DEAE (diethylaminoethanol) Sephadex A-50 gel to remove the coagulation factor IX in the cold-glue plasma; removing impure proteins in the solution by using polyethyleneglycol; carrying out S / D virus inactivation; carrying out anion exchange column chromatography to obtain a purified coagulation factor IX solution; passing through a heparin affinity column for further chromatography to obtain a high-purity coagulation factor IX solution; carrying out ultrafiltration, dialysis and concentration, and adding arginine hydrochloride and glycinate as protective agents; filtering through a 20nm filter element to remove viruses; carrying out freeze-drying; and carrying out dry heat virus inactivation. The protein protective agents are added during the gel adsorption, column chromatography and ultrafiltration dialysis, thereby lowering the activation probability of the FIX product thrombin and enhancing the qualification rate of the product. The technique has high product yield; the FIX specific activity can reach 150 IU / mg or so which is much higher than that of the traditional product; and by performing the three-step virus inactivation, the product is safe and reliable to use.

Description

Technical field

[0001] The invention belongs to the field of biomedicine, and relates to a human coagulation factor IX, specifically a method for preparing high-purity human coagulation factor IX. Background technique

[0002] Human coagulation factor IX (FIX) is one of the thirteen types of coagulation factors in the human body and plays a very important role in the coagulation mechanism of the human body. Blood coagulation disorder caused by the lack of coagulation factor IX is hemophilia B. This disease is a genetic disease. The patient is mainly male, and the incidence is about 1 / 30000 of male live infants. It is spontaneous or traumatic. Bleeding is characterized by bleeding, which is mainly in joints, soft tissues and muscles. Delayed or insufficient treatment can cause various complications, including arthropathy and synovitis associated with repeated joint bleeding, which can lead to joint deformities and dysfunction in severe cases. In addition, more severe bleeding, e...

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