人凝血因子Ⅷ工艺过程效价检测与分析  

Determination and analysis of titer of coagulation factor Ⅷ in manufacturing process

作　　者：李青[1] 刘晓[1] 文圆[1] 窦姿 张安山[1] 张金[1] 何彦林[1] 

机构地区：[1]兰州生物制品研究所有限责任公司甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处：《中国生物制品学杂志》2017年第2期186-191,共6页Chinese Journal of Biologicals

摘　　要：目的测定人凝血因子Ⅷ(human coagulation factorⅧ,FⅧ)生产过程中各关键质控点的FⅧ活性,计算各工序收率。方法采用FⅧ效价测定法(一期法)测定原料血浆、冷沉淀溶解液、化学沉淀后溶液、S/D病毒灭活后溶液、层析纯化洗脱液、超滤后原液、冻干后样品及干热后样品中FⅧ的效价;Lowry法测定各样品的蛋白含量并计算比活;根据各步制品的效价和重量计算各工序FⅧ活性的收率。结果 10批血浆中FⅧ效价平均值为(0.62±0.17)IU/ml,比活平均值(0.011±0.003)IU/mg;FⅧ活性在各步工序中回收率为:离心冷沉淀工序73.78%,化学沉淀工序79.64%,S/D病毒灭活工序93.10%,离子交换层析工序64.96%,超滤工序94.20%,冻干工序90.33%,干热病毒灭活工序79.25%,整个工艺FⅧ的总活性回收率23.96%。按照血浆中FⅧ平均含量0.62 IU/ml计算,每升血浆可产出FⅧ149 IU。7批FⅧ的生产过程中,原液中FⅧ比活较冷沉淀平均提高了170倍。结论获得了由原料血浆到FⅧ成品各关键工序的收率,为生产出高纯度、高质量的FⅧ提供了数据支持。Objective To determinate the titer coagulation factor Ⅷ（FⅧ）in each critical stage of manufacturing process and calculate t he recovery rate. Methods One stage-assay was used to determine the titer of F Ⅷ in material plasma,solution of cryoprecipitate, solution of chemical precipitate, solution after virus inactivation by S / D method, eluate of chromatography, bulk obtained after ultrafiltration, as well as samples taken after lyophilization and virus inactivation by dry heating. The protein content of each sample was determined by Lowry method, based on which the specific activity was calculated. The recovery rate of F Ⅷ activity in each process was calculated according to the titer and weight of product. Results The mean FⅧ titer in 10 batches of plasma pool was（0. 62 ± 0. 17）IU / ml, while the mean specific activity was（0. 011 ± 0. 003） IU / mg. The recovery rates of F Ⅷ activity in the process of centrifugation, chemical precipitation, virus inactivation by S / D method, ion exchange chromatography, ultrafiltration, lyophilization and virus inactivation by dry heating were 73. 78%, 79. 64%, 93. 10%, 64. 96%, 94. 20%, 90. 33% and 79. 25%, respectively,while the total recovery rate in the whole manufacturing process was 23. 96%. Calculated with a mean F Ⅷ content of0. 62 IU / ml, 149 IU FⅧ could be obtained from each liter of plasma. The specific activity of F Ⅷ in the bulk increased by 170 fold in average compared with that in cryprecipitate during production of seven batches of F Ⅷ. Conclusion The recovery rate of FⅧ activity in each process was obtained, which provided a support for the production of F Ⅷ with high purity and high quality.

关 键 词：人凝血因子Ⅷ 效价 比活 工艺过程 收率 

分 类 号：R554[医药卫生—血液循环系统疾病]