Pre-purifying method for cryoprecipitate

Abstract

The invention discloses a pre-purifying method for a cryoprecipitate. The method comprises the following steps that 1, the cryoprecipitate is taken and dissolved; 2, aluminum hydroxide gel dry powder is added, the using amount of the aluminum hydroxide gel dry powder accounts for 5-30%w / w of that of the cryoprecipitate, and stirring is performed for 15-30 min; 3, polyethylene glycol is added until the final concentration of the polyethylene glycol is 3.0-4.0%w / v, the mixture is evenly mixed, the pH is regulated to 6.0-6.5, and stirring is continuously performed for 15-60 min; 4, centrifuging is performed for 10-30 min to obtain supernatant, filtering is performed with a filter membrane, and then the method is completed. According to the pre-purifying method, the purifying effect is good, the FVIII specific activity is increased by 3.69-5.64 times, operation is easy, and the quality is controllable.

Description

technical field [0001] The invention relates to a preparation method of blood products, in particular to a prepurification method of cryoprecipitation. Background technique [0002] Cryoprecipitate is a kind of blood component separated from healthy human plasma. It is a uniform milky white to yellowish viscous precipitate. It is the raw material for large-scale preparation of human coagulation factor VIII (FVIII) concentrated preparations. At present, the commonly used FVIII purification process at home and abroad is to use cryoprecipitation as raw material, to remove most of the impurity proteins after preliminary purification, and to use anionic gel to remove the impurity proteins from the difference in the protein adsorption capacity of the pre-purified feed liquid to obtain high-purity FVIII. finished product. [0003] For the multi-component raw material of cryoprecipitation, the pre-purification process directly affects the quality of the loading liquid (pre-purifica...

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Problems solved by technology

[0003] For cryoprecipitation, a multi-component raw material, the pre-purification process directly affects the quality of the sample feed solution (pre-purification feed solution), which leads to the significant effect of ion exchange chromatography on the purification effect of FVIII and the service life of the gel. difference

[0004] Cryoprecipitation pre-purification methods include aluminum hydroxide gel adsorption, glycine precipitation, polyethylene glycol precipitation and acid precipitation. They have been combined, but the purification effect is not good

[0005] For example, Luo Liang et al., "Preparation of Human High-Purity FVIII Preparation by Ion Exchange Chromatography", West China Pharmaceutical Journal 2000, 15(3): 174-176 disclosed a pre-purification method, specifically, first cryoprecipitated with heparin sodium The specific activity of FVIII is only increased by 1.85 times (0.61 / 0.33, see Table 5 for details), and heparin sodium is required to be purified by dissolving in aqueous solution, followed by adsorption by aluminum hydroxide, acid precipitation and polyethylene glycol precipitation. Pretreatment involves multiple heating and cooling processes, complicated operation and high cost

[0006] Deng Zhihua et al., "Study on Optimization of Extraction Process of Human Coagulation Factor VIII", Jiangxi Medicine, Volume 49, No. 2, February 2014, disclosed a pre-purification method. Specifically, the cryoprecipitate was first dissolved with heparin sodium aqueous solution, and then subjected to hydrogenation. Aluminum adsorption and acid precipitation treatment, using this method to purify, the highest specific activity of FVIII is only increased by 1.9 times, and it needs to be pre-treated with heparin sodium, involving multiple heating and cooling processes, and it needs to prepare aluminum hydroxide gel , complex operation and high cost

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