摘要
目的优化人凝血因子Ⅷ(FⅧ)原液制备工艺,提高FⅧ收得率及纯度。方法采用正交设计优化冷沉淀溶解工艺参数;探讨不同p H对冷沉淀溶解液酸性沉淀的影响,优化得到最佳酸性沉淀条件;通过对离子交换层析工艺的研究,得到离子交换层析的最佳上样流速及柱载量。结果冷沉淀溶解的最佳溶解工艺条件是溶解倍数为4倍、溶解温度为10℃、溶解时间为2 h;溶解液最佳酸性沉淀p H为7.2;离子交换层析的最佳上样流速为1.2 cm·min-1,每毫升凝胶对凝血因子Ⅷ的载量为80.1 IU·m L-1。结论通过对FⅧ原液制备工艺的优化,FⅧ收得率提高了约13%,比活性约提高了30 IU·mg-1。
Objective To improve the recovery rate and purity of human coagulation factor Ⅷ（ F Ⅷ） by optimizing its preparation process. Methods Orthogonal design was used to optimize the parameter of cryoprecipitate dissolving process, to get the best acid precipitation conditions by exploring the effect on the acid precipitates of cryoprecipitate dissolving reagent by different p H values, and to get the best flow rate and column loads of ionexchange chromatography by studying the ion-exchange column chromatograph process. Results The best parameter of cryoprecipitate was that the dissolve ratio was 4, the dissolve temperature was 10 ℃, the dissolve time was 2 h, the p H of the best precipitates was 7.2, the best sample flow rate of ion-exchange column chromatography was 1.2 cm·min^- 1, and the column loads was 80.1 IU·m L^- 1 F Ⅷ gel. Conclusion By optimizing the preparation, the recovery of F Ⅷ has increased about 13%, and the specific activity has increased about 30 IU·mg^- 1.
出处
《中南药学》
CAS
2015年第2期141-143,共3页
Central South Pharmacy
关键词
人凝血因子Ⅷ
冷沉淀
离子交换
层析
human coagulation factor Ⅷ
cryoprecipitate
ionexchange
chromatograph
