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Preparation method of human fibrinogen

A technology of human fibrinogen and raw materials, which is applied in the direction of preparation methods of fibrinogen and peptides, animal/human protein, etc., can solve the problems of complicated production process steps, product purity and biological activity are not very ideal, and improve the comprehensive Utilization ability, biological activity and high purity, and the effect of easier monitoring of quality

Inactive Publication Date: 2016-04-20
TONROL BIOLOGICAL PHARM CO LTD
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  • Claims
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Problems solved by technology

[0002] Human fibrinogen is mainly rich in cryoprecipitate and component I. At present, domestic manufacturers with human fibrinogen production mainly include Hualan Biotechnology, Jiangxi Boya, etc., and most of the manufacturers use components from component I. Extracted by ethanol precipitation twice, the production process steps are complicated, and the purity and biological activity of the final product are not very ideal

Method used

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  • Preparation method of human fibrinogen
  • Preparation method of human fibrinogen

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Experimental program
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Embodiment 1

[0027] Cryoprecipitate was prepared from human plasma by cryogenic centrifugation.

[0028] a. Plasma melting

[0029] Plasma thawing should be carried out in a class 100,000 clean room. Water bath melting method. The water temperature of the water bath melting method should be controlled below 37°C.

[0030] b. Plasma pooling

[0031] Plasma merging should be carried out under a 100,000-level vent. Before merging, plasma bags should be sterilized, that is, soaked in 75% alcohol for more than three minutes, and the outside of the plasma bag should be wiped with clean gauze before merging. When merging plasma, attention should be paid to drip recovery.

[0032] c. Centrifugal separation of cryoprecipitate

[0033] Lower the temperature of the mixed plasma to below 0°C, and perform centrifugation at 3±1 L / unit / min, and control the outlet temperature at 0-4°C.

[0034] Attached to the quality standard of cryoprecipitated raw materials

[0035] 1. The raw material for cryop...

Embodiment 2

[0050] Process for extracting human fibrinogen from cryoprecipitate

[0051] (1) Dissolving the cryoprecipitate: Cut the freshly prepared cryoprecipitate into 1-3 cm blocks, and dissolve it with 3 times the volume of Tris-HCl solution with a concentration of 0.01-0.05mol / L at 24-26°C , adding heparin sodium solution to a final concentration of 0.005-0.065%, adding aluminum hydroxide gel suspension to a final concentration of 5%, stirring for 30 minutes and centrifuging to collect the supernatant.

[0052] (2) Clarification and filtration: perform clarification and filtration with a pretreated filter element with a pore size of 1 μm, and rinse the filter element with a balance solution to 4 times the volume of the cryoprecipitate.

[0053] (3) S / D inactivation: accurately measure the volume of the filtrate, slowly add the prepared S / D solution under stirring at a volume ratio of 1:10, so that the final concentration of polysorbate 80 in the protein solution is 1%, phosphoric ac...

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Abstract

The invention discloses a preparation method of human fibrinogen. The method comprises the following steps that 1, cryoprecipitate serves as a staring material and is dissolved; 2, clarification and filtration are conducted; 3, solvent / detergent (S / D) inactivation is conducted; 4, Q Sepharose fastflow gel adsorption chromatographic processing is conducted; 5, centrifugation is conducted, precipitation is collected, sterilizing, subpackaging, freeze-drying, capping and dry-heating inactivating are conducted, and the product is obtained. The preparation method of the human fibrinogen is simple, easy to operate, low in cost and capable of increasing the comprehensive utilization ratio of blood plasma, and the obtained product is high in purity, activity and safety.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and in particular relates to a preparation method of fibrinogen. Background technique [0002] Human fibrinogen is mainly rich in cryoprecipitate and component I. At present, domestic manufacturers with human fibrinogen production mainly include a few such as Hualan Biotechnology and Jiangxi Boya, and most of the manufacturers use components from component I. Extracted by ethanol precipitation twice, the production process steps are complicated, and the purity and biological activity of the final product are not very ideal. [0003] Human fibrinogen is a blood coagulation factor, and its clinical application is mainly to treat acquired fibrinogen reduction: severe liver injury; liver cirrhosis; disseminated intravascular coagulation; postpartum hemorrhage and fibrosis caused by major surgery, trauma or internal bleeding, etc. Coagulation disorders caused by protein deficiency. Human fibrinogen...

Claims

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Application Information

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IPC IPC(8): C07K14/75C07K1/16
CPCC07K14/75
Inventor 胡辉恒邓昆魏成
Owner TONROL BIOLOGICAL PHARM CO LTD
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