不同直径层析柱纯化人凝血因子Ⅷ效果的比较  

Comparison of purification effects of human coagulation factorⅧby chromatography columns with different diameters

作　　者：刘环 曹璟 吴鹏 李勇 许周敏 江砚芳 Liu Huan;Cao Jing;Wu Peng;Li Yong;Xu Zhoumin;Jiang Yanfang(Blood Preparation Department,Sinopharm Shanghai Plasma-derived Biotherapies Co.,Ltd.,Shanghai 200051,China;Blood Preparation Department,Sinopharm Kunming Plasma-derived Biotherapies Co.,Ltd.,Kunming 650212,China)

机构地区：[1]国药集团上海血液制品有限公司血液制剂室,上海200051 [2]国药集团昆明血液制品有限公司血液制剂室,昆明650212

出　　处：《国际生物制品学杂志》2024年第2期94-98,共5页International Journal of Biologicals

摘　　要：目的通过比较RJ300型和GE450型层析柱对人凝血因子Ⅷ(human coagulation factor Ⅷ, F Ⅷ)的纯化效果, 探究GE450型层析柱纯化FⅧ的适用性。方法分别采用2种层析柱各制备3批FⅧ制品, 按照中国药典2020年版三部的要求对制品进行检测, 比较2种层析柱制备的FⅧ原液的回收率、比活性、洗脱峰的峰形差异、杂蛋白去除效果以及成品的检定结果, 并进行产品稳定性考察。结果 RJ300型和GE450型层析柱纯化后的FⅧ原液回收率分别为(63.8±4.8)%和(61.0±4.3)%, 相对标准偏差分别为7.5%和7.0%, 原液比活性分别为(100.7±5.8)和(114.0±12.2)国际单位(international unit, IU)/mg, 2种层析柱洗脱峰的峰形差异无统计学意义(F=0.42, P>0.05), 2种层析柱均能有效去除杂蛋白, 成品检定结果均合格, 成品比活性分别为(38.6±5.5)和(39.4±6.2) IU/mg。2种层析柱制备的FⅧ的稳定性研究结果均符合中国药典规定。结论 GE450型层析柱能实现较好的FⅧ纯化效果, 成品安全有效、稳定性良好, 适合商业规模化生产。Objective To explore the applicability of GE450 chromatography column for purification of human coagulation factorⅧ(FⅧ)by comparing the purification effects of RJ300 and GE450 chromatography columns.Methods Three batches of FⅧproducts each were prepared by two types of chromatography columns respectively,and the products were detected according to the requirements of Chinese pharmacopoeia 2020 edition volumeⅢ.The recovery rate,specific activity,difference of elution peak,removal effect of impurities for FⅧbulk and detection results of the final product prepared by two chromatography columns were compared.The stability of the products was investigated.Results The recovery rates of FⅧbulk purified by RJ300 and GE450 chromatography columns were(63.8±4.8)%and(61.0±4.3)%,respectively,the relative standard deviation were 7.5%and 7.0%,respectively,and the specific activity of bulk were(100.7±5.8)and(114.0±12.2)international unit(IU)/mg,respectively.There was no statistically significant difference(F=0.42,P>0.05)in elution peaks between the two columns,and both columns effectively removed impurities.The results of the final products were qualified,and the specific activity of the final products were(38.6±5.5)and(39.4±6.2)IU/mg,respectively.The results of stability study for FⅧprepared by both types of chromatography columns met the requirements of Chinese pharmacopoeia.Conclusion GE450 chromatography column achieves good purification effect for FⅧ,and the final product is safe,effective and stable,thus is suitable for commercial scale production.

关 键 词：人凝血因子Ⅷ 层析柱 离子交换层析 纯化 回收率 比活性 稳定性 

分 类 号：TQ464[化学工程—制药化工]