A preparing process for instant precipitate-free freeze-dried human fibrinogen

Abstract

The invention relates to the field of preparing processes for human fibrinogen, and particularly relates to a preparing process for instant precipitate-free freeze-dried human fibrinogen. The process includes five steps, namely the method includes a first step of component I precipitate dissolving and filtering; a second step of S / D virus inactivation; a third step of twice low-temperature ethanol precipitation rectification; a fourth step of freeze-drying; and a fifth step of virus inactivation in a dry heat manner. Through the improved fibrinogen production process, the fibrinogen which is instant, precipitate-free, opalescence-free and free of protein particles can be prepared. The process can prepare the human fibrinogen used for traditional intravenous injection, and can prepare high-concentration human fibrinogen compatible in human fibrous protein adhesives.

Description

Technical field [0001] The invention belongs to the field of biopharmaceuticals, and relates to a preparation process of blood products, in particular, a preparation process of freeze-dried human fibrinogen with no precipitation. Background technique [0002] Human fibrinogen (fibrinogen or Fg for short) is a macromolecular glycoprotein containing 2964 amino acids. It is a protein with coagulation function synthesized by the liver. It has the largest content of thirteen coagulation factors in the human body. A coagulation factor, also known as coagulation factor I. Human fibrinogen is a blood product that is frequently used clinically, and it is also an indispensable emergency medicine for hemostasis on the battlefield. However, fibrinogen has been in a clinically in short supply. Especially in recent years, human fibrin adhesive, a surgical blood product with highly effective hemostasis function, has become more and more popular among doctors and patients. As one of the compati...

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Problems solved by technology

[0004] The fibril preparation process using cold glue as raw material, because the raw material contains very little Fg, after a series of operation steps, layer by layer loss, the final yield is very limited, lacking the value of commercial production; while cold glue and components I co-precipitation is the fibroblast production technique of raw material, also there is following problem, at first, contain 8% ethanol in the precipitation, for the stability of the human coagulation factor VIII in the precipitation, be a very unfavorable factor, the storage of precipitation It must be very careful, the storage time should not be too long, and special attention must be paid to the denaturation and damage of ethanol to human blood coagulation factor VIII when feeding materials, so the existence of ethanol in raw materials has undoubtedly proposed harsh requirements for the production process of human blood coagulation factor VIII; secondly, due to The presence of a large amount of human coagulation factor VIII in the precipitate also brings hidden dangers to the production of fibrils, because fibrils are easily activated and lead to production failure. The reasons for fibril activation are complicated, but thrombin is the most direct reason. Thrombin is prothrombin in Ca 2+ Ions and other blood coagulation factors, including blood coagulation factor VIII, are activated through a series of complicated chain reactions. Therefore, the existence of human blood coagulation factor VIII is also an unfavorable factor during the preparation of fibrils. It is completely removed in the initial process; in addition, from the perspective of the preparation process of fibrils, some processes are complicated, especially the dissolution and precipitation are repeated, which leads to frequent heating, cooling, adding ethanol, acid-base and other operations. The operation will cause damage to the protein to varying degrees. Once there is a drastic change in the operation, it is likely to cause opalescence in the final product and even a long time for precipitation and reconstitution.

In addition, the excessive pursuit of the purity of fibroblasts is also the reason why some processes are complicated and the production cycle is too long. Many practices have proved that excessive product purity is not good for stability, and it is easy to cause fibrils to separate out.

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