Method for preparing human prothrombin complex from plasma

A technology of human prothrombin and complexes, which is applied in the field of preparation of human prothrombin complexes, can solve the problems of prothrombin activation, blood coagulation reaction, gel leakage, etc., to reduce the mixing of foreign proteins, reduce pollution, The effect of increasing material loss

Active Publication Date: 2017-05-17
广东双林生物制药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The batch adsorption process is an open operation, the operation is cumbersome, and it is easy to cause pollution, cross-contamination, gel leakage and other problems
Domestic human prothrombin complexes are also reported

Method used

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  • Method for preparing human prothrombin complex from plasma

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Embodiment 1

[0027] Removal of plasma cryoprecipitate: using fresh frozen human plasma as raw material, the cryoprecipitate in the plasma is removed through the processes of thawing, mixing, and continuous centrifugation to obtain 2 kg of plasma centrifugation supernatant without cryoprecipitation;

[0028] Gel batch adsorption: The plasma temperature is controlled at 10°C. According to 1.0g dry gel / L plasma, add the balanced DEAE A-50 gel to the cryoprecipitated plasma, stir and absorb for 45min, and then turn off the stirring. The plasma after gel adsorption is filtered with a self-flushing filter, the absorbed A-50 gel is collected, and the plasma filtrate is incorporated into the plasma tank;

[0029] Column packing and elution: Pack the adsorbed A-50 gel in a fixed bed chromatography column, and use a peristaltic pump to pump washing liquid into the packed chromatography column for online flushing. The flow rate of the washing liquid is 35cm / h, and the flushing time About 60min. The ...

Embodiment 2

[0034] Removal of plasma cryoprecipitate: using fresh frozen human plasma as raw material, the cryoprecipitate in the plasma is removed through the processes of thawing, mixing, and continuous centrifugation to obtain 2 kg of plasma centrifugation supernatant without cryoprecipitation;

[0035] Gel batch adsorption: plasma temperature is controlled at 15°C, according to 1.5g dry gel / L plasma, add balanced DEAE A-50 gel to the cryoprecipitated plasma, stir and absorb for 45 minutes, turn off the stirring, and let it settle for 45 minutes . The plasma after gel adsorption is filtered with a self-flushing filter, the absorbed A-50 gel is collected, and the plasma filtrate is incorporated into the plasma tank;

[0036] Column packing and elution: Pack the adsorbed A-50 gel in a fixed-bed chromatography column, and use a peristaltic pump to pump washing liquid into the packed chromatography column for online flushing. The flow rate of the washing liquid is 40cm / h, and the flushing ti...

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Abstract

The invention discloses a method for preparing a human prothrombin complex from plasma. The method comprises: directly adsorbing the human prothrombin complex from the plasma by using DEAE A-50 gel, filling the adsorbed A-50 gel into a chromatographic column of a fixed bed, pumping eluant into the filled chromatographic column by using a peristaltic pump for online washing and elution, performing S/D inactivation on the eluant, and performing secondary chromatographic purification by using the chromatographic column of the fixed bed to obtain a high-purity human prothrombin complex product. According to the method, elution flow and speed can be accurately controlled by online elution after filling the gel into the column, the problems of contamination, cross contamination, gel leakage and the like caused by an open operation are reduced, an obtained product is high in purity, an IX factor titer can exceed 27IU/ml, and the IX factor specific activity exceeds 0.8IU/mg protein. Meanwhile, a self-flushing type filter with a pressure difference controller and an automatic solid matter stripping system is adopted, so the pressure difference in the whole filtering process is stable and controllable, gel particles can be well protected, broken colloidal particles flowing into a plasma tank in the filtering process are remarkably reduced, and gel losses in a production process can be reduced by above 20%.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals and blood products, and in particular relates to a method for preparing human prothrombin complex from blood plasma in the preparation of blood products. Background technique [0002] Human prothrombin complex concentrate (PCC) is derived from healthy human plasma and is a traditional plasma protein product. In addition to being used for the treatment of hemophilia B, it can also be used for the treatment of hemophilia caused by vitamin K deficiency or liver disease. With the gradual recognition of clinicians, the use of PCC is also increasing year by year. At present, there are not many manufacturers of PCC in our country, and the raw material plasma is in short supply, so there is a serious shortage of PCC. Therefore, its production and preparation have attracted attention from all sides. [0003] At present, the classic process for preparing human prothrombin complex is the batch ad...

Claims

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Application Information

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IPC IPC(8): C12N9/74
CPCC12N9/6429C12Y304/21005
Inventor 蒋桂香朱光祖胡川骆燕容罗观文洪好武梁栋立
Owner 广东双林生物制药有限公司
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