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Method for purifying human coagulation factor VII from human plasma

A technology of human blood coagulation factor and human plasma, which is applied in the field of bioengineering, can solve the problems of decreased titer of blood coagulation factor VII, achieve high specific activity, reduce pollution, and improve utilization rate

Active Publication Date: 2020-08-07
HUALAN BIOLOGICAL ENG CHONGQING
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Problems solved by technology

[0003] Human blood coagulation factor VII is commonly used clinically for hemophiliacs and traumatic bleeding patients. With the gradual expansion of medical insurance coverage, the dosage of blood coagulation factor VII drugs will continue to increase. Human blood coagulation factor VII is relatively stable, but in the preparation process, it also includes virus A method of inactivation or removal that results in a decrease in the titer of coagulation factor VII

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  • Method for purifying human coagulation factor VII from human plasma

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Embodiment 1

[0021] A method for purifying human blood coagulation factor VII from human plasma, comprising the following steps:

[0022] S1-de-cold plasma:

[0023] Thaw the fresh frozen plasma, and continuously centrifuge it at a temperature of 0-3°C to remove the cold glue. cold-depleted plasma;

[0024] S2- Preparation before the first chromatography:

[0025] UniGel-80Q ion exchange packing chromatography column preparation, equilibrate UniGel-80Q gel packing with a buffer containing 0.01mol / L~0.02mol / L sodium citrate and 0.1mol / L~0.15mol / L sodium chloride, The pH of the buffer solution is 6.80-7.80, after treatment, wait for sample loading.

[0026] S3-first anion exchange chromatography:

[0027] Adjust the plasma conductivity to 10-13mS / cm, the pH value to 7.00-8.00, and the protein content to 45g / L±5g / L, and load the adjusted gel-free plasma on the UniGel-80Q gel chromatography column to make the blood coagulation factor The target protein of VII is adsorbed in the UniGel-80Q...

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Abstract

The invention discloses a method for purifying a human coagulation factor VII from human plasma. The method comprises the following steps: (1) removing cold gel plasma, (2) performing anion exchange chromatography for the first time, (3) filtering the eluent, and adjusting the conductivity and pH value of the product, and (4) carrying out ion exchange chromatography for the second time. Accordingto the method, the human coagulation factor VII can be separated from the cold-gel-free plasma by adopting sodium glutamate as an eluent through first ion exchange chromatography, and the human coagulation factor VII can be further purified through two times of chromatography, so that the utilization rate of the plasma is greatly improved, and the human coagulation factor VII can achieve relatively high specific activity.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for purifying human blood coagulation factor VII from human blood plasma. Background technique [0002] Coagulation factors are protein components involved in the blood coagulation process. There are more than a dozen coagulation factors involved in blood coagulation in plasma. Among them, human coagulation factor VII is the initial factor that triggers the coagulation cascade reaction in the extrinsic coagulation pathway. Hepatocyte synthesis of vitamin K-dependent zymogens with serine protease hydrolysis. In addition to the treatment of coagulation factor VII deficiency, human coagulation factor VII has also been widely studied and applied in the treatment of hemophilia patients. At present, the treatment of hemophilia patients is facing severe challenges, because up to 20% of patients developed coagulation factor inhibitors to coagulation factor VIII or coagul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64
CPCC12N9/6451C12Y304/21021
Inventor 肖岚张海梦李凯旋滕世超张宝献张建璀刘余江
Owner HUALAN BIOLOGICAL ENG CHONGQING
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