Preparation method of human thrombinogen compound

Abstract

The invention belongs to the technical fields of bio-pharmaceuticals and blood products, and particularly relates to a preparation method of a human thrombinogen compound. According to the human thrombinogen compound prepared by the method, an excellent production process is obtained by improving and researching adsorbing balance liquid, subsequent washing liquid and eluent and a freeze-drying process, four blood coagulation factors that the PCC product contain have high active ratio, the yield of each component is high, the IX specific activity is up to 3.13 IU / mg, the specific activity is higher than that of the pharmacopeia standard, the thrombosis probability is effectively reduced and activation of the blood coagulation factors is reduced.

Description

technical field

[0001] The invention belongs to the technical field of biopharmaceuticals and blood products, and in particular relates to a preparation method of human prothrombin complex. Background technique

[0002] Human prothrombin complex concentrates (PCC) is a plasma protein preparation containing four vitamin K-dependent coagulation factors, coagulation factors II, VII, IX, and X, which can promote blood coagulation. Clinically, PCC is mainly used to treat hemophilia B and liver disease, as well as bleeding symptoms caused by low coagulation factor levels caused by vitamin K deficiency. In addition, it has also been affirmed in the treatment of bleeding symptoms caused by FVIII inhibitors in hemophilia A patients who have been infused with FVIII for a long time. In foreign countries, PCC is also widely used to treat bleeding symptoms caused by overdose of vitamin K antagonists as anticoagulant drugs. In recent years, with the recognition of clinicians, the demand...

Examples

Embodiment 1

[0037] A preparation method of human prothrombin complex, comprising the following preparation steps:

[0038] 1) Centrifugation: Using fresh frozen human blood as raw material, the cryoprecipitate in the plasma is removed by melting, mixing, and centrifugation. After centrifugation, the plasma supernatant is clarified and filtered with a PALL filter equipped with a 0.45um filter element to obtain plasma. supernatant;

[0039] 2) Adsorption: adjust the pH of the plasma supernatant, add DEAE Sephadex A-50 gel, 1.0°C, stir and adsorb for 30 minutes, then filter;

[0040] 3) Washing and elution: DEAE Sephadex A-50 gel is installed in the chromatography column, and the impurity protein is washed on-line with the washing liquid, 50cm / h, flushing 5 column volumes, and the adsorption II in the gel is eluted with the eluent , VII, IX, X factors and a small amount of protein, the elution speed is 30cm / hn, stop elution when the detection value of the online detector is lower than 500Au...

Embodiment 2-8

[0058] Table 1 Example 2-8 adsorption conditions

[0059]

[0060]

[0061] Other preparation steps of Examples 2-8 are the same as Example 1.

Embodiment 9-15

[0063] Table 2 embodiment 9-15 washing conditions

[0064]

[0065] Other preparation steps of Examples 9-15 are the same as Example 1.

technical field

[0001] The invention belongs to the technical field of biopharmaceuticals and blood products, and in particular relates to a preparation method of human prothrombin complex. Background technique

[0002] Human prothrombin complex concentrates (PCC) is a plasma protein preparation containing four vitamin K-dependent coagulation factors, coagulation factors II, VII, IX, and X, which can promote blood coagulation. Clinically, PCC is mainly used to treat hemophilia B and liver disease, as well as bleeding symptoms caused by low coagulation factor levels caused by vitamin K deficiency. In addition, it has also been affirmed in the treatment of bleeding symptoms caused by FVIII inhibitors in hemophilia A patients who have been infused with FVIII for a long time. In foreign countries, PCC is also widely used to treat bleeding symptoms caused by overdose of vitamin K antagonists as anticoagulant drugs. In recent years, with the recognition of clinicians, the demand...

Examples

Embodiment 1

[0037] A preparation method of human prothrombin complex, comprising the following preparation steps:

[0038] 1) Centrifugation: Using fresh frozen human blood as raw material, the cryoprecipitate in the plasma is removed by melting, mixing, and centrifugation. After centrifugation, the plasma supernatant is clarified and filtered with a PALL filter equipped with a 0.45um filter element to obtain plasma. supernatant;

[0039] 2) Adsorption: adjust the pH of the plasma supernatant, add DEAE Sephadex A-50 gel, 1.0°C, stir and adsorb for 30 minutes, then filter;

[0040] 3) Washing and elution: DEAE Sephadex A-50 gel is installed in the chromatography column, and the impurity protein is washed on-line with the washing liquid, 50cm / h, flushing 5 column volumes, and the adsorption II in the gel is eluted with the eluent , VII, IX, X factors and a small amount of protein, the elution speed is 30cm / hn, stop elution when the detection value of the online detector is lower than 500Au...

Embodiment 2-8

[0058] Table 1 Example 2-8 adsorption conditions

[0059]

[0060]

[0061] Other preparation steps of Examples 2-8 are the same as Example 1.

Embodiment 9-15

[0063] Table 2 embodiment 9-15 washing conditions

[0064]

[0065] Other preparation steps of Examples 9-15 are the same as Example 1.

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