A preparation process for extracting human fibrinogen from the waste material of coagulation factor ⅷ extracted by cryoprecipitation

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...

AI Technical Summary

Problems solved by technology

Fibrinogen is readily activated, especially at Ca 2+ In the presence of thrombin, thrombin activates the otherwise water-soluble fibrinogen into fibrin monomers, which are polymerized and produced at Ca 2+ In the presence of water-insoluble fibrin is formed, and the fibrin is then twisted into agglomerates to form fibrin glue. Therefore, it is easy to cause difficulty in filtration and long reconstitution time of the product during the process of preparation

[0009] It can be seen that there are still some problems in the existing human fibrinogen: (1) The purity is not high, and the product contains too many impurities, which is prone to adverse reactions such as disseminated intravascular coagulation, rash, tachycardia, and fever in clinical use; (2) The solubility is low, and the reconstitution time is too long during use, which is inconvenient to use, especially in emergency situations

The reconstitution time of most products is about 20 minutes, and some even reach about 30 minutes; (3) The yield is not high, especially for the extraction of human fibrinogen from cryoprecipitate

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