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Preparation process of fibrinogen

A fibrinogen and preparation process technology, applied in the field of blood products, can solve the problems of increasing process time and equipment cost, the influence of lifespan on chromatographic effects, and increasing process risks, so as to ensure purity and recovery rate, shorten process time, The effect of simple process

Pending Publication Date: 2022-05-10
新疆德源生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, in the "Method for Extracting Human Fibrinogen from Component I by Column Chromatography" published by Chinese patent CN102212129B, the chromatographic column needs to be pre-balanced, and after the chromatography, it needs 3 times the volume of the column bed to wash and merge, which increases significantly. Process time and equipment cost; while the life of the gel in gel adsorption has a significant impact on the chromatography effect, and will increase the process risk
[0007] Therefore, the current domestic production process of human fibrinogen has problems such as high cost, long production cycle, and low yield.

Method used

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  • Preparation process of fibrinogen
  • Preparation process of fibrinogen
  • Preparation process of fibrinogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation process 1 of embodiment 1 fibrinogen

[0041] (1) Preparation of component I: add A-50 gel with 2.0% plasma weight to the cryoprecipitate supernatant, stir and absorb for 60 minutes, let stand for more than 40 minutes to separate, and collect the eluate for prothrombin complex production; collect the flow Make component I through the liquid; add 15°C, 0.9% sodium chloride solution to the flow-through liquid, adjust the pH value of the flow-through liquid to 7.15 with pH4.0 acetate buffer, add -15°C at 0°C, 95% ethanol to a final concentration of 8% v / v, control the temperature at -2.5°C, continue stirring for 2 hours, control the temperature at -3°C, feed the liquid at 50L / min, rotate at 4800 rpm, and centrifuge to obtain Component I precipitates.

[0042] (2) The first step of impurity removal: Precipitate the component I prepared in step (1), chop it up, add 5 times the volume of dissolving solution A to dissolve, stir for 1 hour to prevent foam gener...

Embodiment 2

[0050] The preparation process 2 of embodiment 2 fibrinogen

[0051] (1) Preparation of component I: add A-50 gel with 3.0% plasma weight to the cryoprecipitate supernatant, stir and absorb for 60 minutes, let stand for more than 40 minutes for separation, and collect the eluate for prothrombin complex production; collect the flow Make component I through the liquid; add 0°C, 0.9% sodium chloride solution to the flow-through liquid, adjust the pH value of the flow-through liquid to 7.20 with pH4.0 acetate buffer, add -20°C at 0°C, 95% ethanol to a final concentration of 8% v / v, control the temperature at -2.0°C, continue stirring for 2 hours, control the temperature at -3°C, feed the liquid at 50L / min, rotate at 5200 rpm, and centrifuge to obtain Component I precipitates.

[0052] (2) The first step of impurity removal: Precipitate the component I prepared in step (4), chop it up, put it into 8 times solution A to dissolve, stir to dissolve, stir for 1.5h, prevent foam genera...

Embodiment 3

[0060] The preparation process 3 of embodiment 3 fibrinogen

[0061] (1) Preparation of component I: add A-50 gel with 4.5% plasma weight to the cryoprecipitate supernatant, stir and adsorb for 60 minutes, let stand for more than 40 minutes to separate, and collect the eluate to make prothrombin complex; collect the flow Make component I through the solution; add 10°C, 0.9% sodium chloride solution to the flow-through solution, adjust the pH value of the flow-through solution to 7.30 with pH4.0 acetate buffer, add -10°C at 0°C, 95% ethanol to a final concentration of 8% v / v, control the temperature at -2.5°C, continue to stir for 2 hours, control the temperature at 0°C, feed the liquid at 50L / min, rotate at 5000 rpm, and centrifuge to obtain the composition Part I precipitation.

[0062] (2) The first step of impurity removal: Precipitation of component I prepared in step (1), chopping, adding solution A of 7 times the weight of precipitation, stirring to dissolve, stirring f...

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Abstract

The invention relates to a preparation process of fibrinogen. The preparation process comprises a two-step impurity removal process, a synchronous virus inactivation process by combining an inactivator with a pasteurization method and a two-step purification process. The two-step impurity removal process improves the yield of the crude extract of the product; in the synchronous virus inactivation process by combining the inactivator with the pasteurization method, the influence of high temperature on the quality of the fibrinogen product in the pasteurization process is overcome by optimizing the formula of the inactivator, and the duration of the inactivation process is shortened; the use amount of the inactivating agent is reduced, and the safety of the product is improved; the two-step purification process improves the quality attribute of the product. Therefore, on the basis of comprehensively considering the high efficiency and necessity of the fibrinogen preparation process, the time of the inactivation process is shortened, the extraction process is simplified, the production cost is reduced, meanwhile, the yield is improved, the purity of the obtained fibrinogen product is higher than 90%, the coagulation activity is 16-20 s, the redissolution time is shorter than 12 min, and the yield reaches up to 2461 bottles per ton of plasma.

Description

technical field [0001] The invention belongs to the field of blood products, and in particular relates to a preparation process of fibrinogen. Background technique [0002] Human fibrinogen (Fibrinogen) is human coagulation factor Ⅰ, one of the main components of plasma protein, with a molecular weight of about 340,000 and an isoelectric point of 5.5. It can convert fibrinogen into insoluble Formed components such as fibrin and blood cells in the network coagulate into blood clots to achieve the purpose of hemostasis. Human fibrinogen products can rapidly increase the content of fibrinogen in the blood, and are used to treat postpartum hemorrhage and coagulation disorders caused by fibrinogen deficiency caused by major surgery, trauma or internal bleeding. Fibrinogen is the first clotting factor to reach bed low levels during major bleeding and replacement therapy with red blood cell concentrates. Thus, low perioperative fibrinogen levels have been shown to be associated w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/75C07K1/14C07K1/36C07K1/30C07K1/34
CPCC07K14/75
Inventor 吕献忠苏峰崔婷婷孙书秒王雅琴陈远洋
Owner 新疆德源生物工程有限公司
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