Preparation process of lyophilized human thrombin

Abstract

The invention discloses a preparation process of lyophilized human thrombin. The preparation process is characterized by comprising the following steps: preparing cooled precipitated blood plasma intoa human prothrombin, activating the human prothrombin to obtain a human thrombin crude solution, preparing human thrombin from the human thrombin crude solution, wherein the activation conditions ofthe human prothrombin are that: 0.3mol/L of calcium chloride solution is added in an amount of one tenth of an original human prothrombin solution, so that the calcium chloride final concentration is0.025 to 0.03mol/L, then soybean lecithin or egg yolk lecithin is added, so that calcium chloride with the concentration of 0.1 to 0.2 percent is taken as an activating agent; slowly stirring the solution, and activating at the temperature of 20 to 25 DEG C for 1 to 2 hours to obtain a human thrombin solution. The prepared human thrombin is 400 to 700IU/ml in titer, is over 1,000IU/mg proteins inspecific activity, and is over 20IU/ml in yield. The preparation process has the advantages of good purifying effect, high yield and easiness in operation, and the situations of poor activation effect, low coversion rate, low yield, over long activation time, turbidity after re-dissolving, lowering of the titer after long-time storage in the prothrombin are avoided effectively.

Description

technical field [0001] The invention relates to a preparation process of human thrombin and belongs to the field of biopharmaceuticals. Background technique [0002] As early as the end of the 19th century, thrombin was discovered and named by a Scottish physiologist. It was not until thrombin was purified and sequenced in 1951 that people's research and understanding of thrombin became more and more in-depth. The physiological function of thrombin is related to the activity of its serine protease, which can cleave fibrinogen to form fibrin clot, thereby promoting the formation of blood clot. [0003] Thrombin is a highly specific serine proteolytic enzyme that is activated by prothrombin and is a natural component in the body's blood coagulation system. Thrombin is composed of 308 amino acid residues and has a molecular mass of 36KD. It is a double-chain molecule, containing A chain and B chain, and the two chains are connected by disulfide bonds. The A chain contains 49 ...

AI Technical Summary

Problems solved by technology

[0007] According to the above-mentioned existing preparation methods of thrombin, all are various combinations of prothrombin extraction, activation, and thrombin purification methods, but there may still be some problems in the existing preparation methods, for example, thrombin is very unstable, Thrombin titer reduction and protein denaturation are likely to occur in the state; if the freeze-dried product is not p...

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