Preparation process for extracting human fibrinogens from waste for extracting cryoprecipitated blood coagulation factor VIII

Abstract

The invention discloses a preparation process for extracting human fibrinogens from waste for extracting cryoprecipitated blood coagulation factor VIII. The preparation process is characterized by comprising the steps of cryoprecipitation dissolution, 2% aluminium hydroxide gel absorption, ion strength adjustment, series connection filtering, S / D viral inactivation, ion-exchange chromatography, EDTA Ca2+ removal, glycine precipitation, primary low-temperature ethanol precipitation, AT-III thrombin inhibition, secondary low-temperature ethanol precipitation, nanofilm filtering and dry-heat inactivation. For ensuring the safety, a nanofilm ia added to filter virus except S / D and dry-heat inactivation. By means of an added AT-III inactivated thrombin and EDTA Ca2+ removal process, fibrinogen in the production process is effectively prevented from being activated into fibrous protein. The glycine precipitation is utilized to remove fibrous protein monomers and polymers in products so as obtain high-purity human fibrinogen. The preparation products are safe and reliable, redissolution time is short, the clinic first-aid demand is met, and meanwhile the preparation process has important significance on indirect saving of scarce plasma resources.

Description

technical field

[0001] The invention relates to a preparation process for extracting human fibrinogen from the waste material of coagulation factor VIII extracted by cryoprecipitation, which belongs to the field of biopharmaceuticals. Background technique

[0002] Fibrinogen (Fg), synthesized and secreted by liver cells, is an important coagulation factor in the body's hemostatic physiology. The storage stability is good, but the thermal stability is poor, and irreversible precipitation can be formed at 56°C. The concentration of Fg in normal human plasma is 2.4-4.0g / L.

[0003] Fg is an acute phase reaction protein, and its abnormal blood content is a risk factor for ischemic cardiovascular and cerebrovascular diseases and other diseases. The increase of Fg is often a non-specific reaction of the body, which is common in infections such as toxemia, pneumonia, cholecystitis, tuberculosis, and aseptic inflammations such as nephrotic syndrome, rheumatic fever, malignant tumor...

Examples

Embodiment 1

[0043] Embodiment 1: Taking 20,000 liters of plasma as an example, the specific preparation process is as follows:

[0044] (1) During the quarantine period, after receiving the plasma of qualified individuals, wipe the surface of the plasma bag with 75% ethanol, rinse it with water for injection, merge it into a slurry tank, and melt it with circulating water below 30-35°C. The temperature of the plasma should not be higher than 4°C; after melting, centrifuge, control the liquid temperature at 0-4°C, and collect 136.9kg of cryoprecipitate;

[0045] (2) Add the cryoprecipitate obtained in step (1) into 3IU / ml heparin sodium solution, stir until the cryoprecipitate is completely dissolved, and control the temperature of the circulating water at 20-28°C; start centrifugation, collect the supernatant, Weighed 507.8kg;

[0046] (3) Adjust the pH of the supernatant obtained in step (2) to 6.6-7.2 with 0.5mol / L HCL; add 2% aluminum hydroxide gel, stir; start centrifugation, collect...

technical field

[0001] The invention relates to a preparation process for extracting human fibrinogen from the waste material of coagulation factor VIII extracted by cryoprecipitation, which belongs to the field of biopharmaceuticals. Background technique

[0002] Fibrinogen (Fg), synthesized and secreted by liver cells, is an important coagulation factor in the body's hemostatic physiology. The storage stability is good, but the thermal stability is poor, and irreversible precipitation can be formed at 56°C. The concentration of Fg in normal human plasma is 2.4-4.0g / L.

[0003] Fg is an acute phase reaction protein, and its abnormal blood content is a risk factor for ischemic cardiovascular and cerebrovascular diseases and other diseases. The increase of Fg is often a non-specific reaction of the body, which is common in infections such as toxemia, pneumonia, cholecystitis, tuberculosis, and aseptic inflammations such as nephrotic syndrome, rheumatic fever, malignant tumor...

Examples

Embodiment 1

[0043] Embodiment 1: Taking 20,000 liters of plasma as an example, the specific preparation process is as follows:

[0044] (1) During the quarantine period, after receiving the plasma of qualified individuals, wipe the surface of the plasma bag with 75% ethanol, rinse it with water for injection, merge it into a slurry tank, and melt it with circulating water below 30-35°C. The temperature of the plasma should not be higher than 4°C; after melting, centrifuge, control the liquid temperature at 0-4°C, and collect 136.9kg of cryoprecipitate;

[0045] (2) Add the cryoprecipitate obtained in step (1) into 3IU / ml heparin sodium solution, stir until the cryoprecipitate is completely dissolved, and control the temperature of the circulating water at 20-28°C; start centrifugation, collect the supernatant, Weighed 507.8kg;

[0046] (3) Adjust the pH of the supernatant obtained in step (2) to 6.6-7.2 with 0.5mol / L HCL; add 2% aluminum hydroxide gel, stir; start centrifugation, collect...

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