注射用重组人凝血因子Ⅶa效价检测方法的建立  

Establishment of a method for detecting the potency of recombinant human coagulation factor Ⅶa for injection

作　　者：吴蓉 王俪鲆 郎锦烨 朱玥 周静 刘珣 倪静 周顺波 丁亚凌 WU Rong;WANG Liping;LANG Jinye;ZHU Yue;ZHOU Jing;LIU Xun;NI Jing;ZHOU Shunbo;DING Yaling(Chengdu Rongsheng Pharmaceutical Co.,Ltd,Chengdu 610000,China)

机构地区：[1]成都蓉生药业有限责任公司,四川成都610000

出　　处：《中国输血杂志》2025年第3期415-420,共6页Chinese Journal of Blood Transfusion

基　　金：药品监管科学全国重点实验室课题(2024SKLDRS0207)。

摘　　要：目的 建立适用于注射用重组人凝血因子Ⅶa效价检测的一种方法。方法 通过在样品杯中加入样品和乏Ⅶ因子血浆,用凝血酶原时间测定试剂(PT试剂)激活反应,通过样品杯中磁珠摆动振幅的变化判定样品的凝固时间,凝固时间的对数与人凝血因子Ⅶa效价的对数成反比。结果 在本试验条件下,通过加标回收考察方法学的专属性,回收率均在90.0%~110.0%之间。在0.125~1.000 IU/mL范围内,标准品、样品的效价与凝固时间之间呈现良好的线性响应,相关系数r均>0.99。准确性、重复性:原液检测的各浓度回收率分别为101.0%、100.0%、112.0%,RSD分别为2.6%、4.0%、0.0%;成品检测的各浓度回收率分别为104.0%、94.7%、112.0%,RSD分别为1.9%、2.4%、0.0%。中间精密度:RSD_(12)次分别为4.5%、3.7%。样品经稀释液处理后,常温放置6 h内检测,仍具有较稳定的生物学活性。结论 该检测方法的结果准确稳定、操作简便、自动化程度高,适用于注射用重组人凝血因子Ⅶa效价的测定。Objective To establish a method for detecting the potency of recombinant human coagulation factorⅦa for injection.Methods By adding the sample and factorⅦdeficient plasma to the sample cup and activating the reaction with prothrombin time assay reagent(PT reagent),the coagulation time of the sample was determined by the change in magnetic bead swing amplitude in the sample cup.The logarithm of coagulation time was inversely proportional to the logarithm of hu-man factorⅦa potency.Results Under the experimental conditions,the specificity of the methodology was evaluated through spiked recovery,and the recovery rates ranged from 90.0%to 110.0%.Within the range from 0.125 to 1.000 IU/mL,there was a good linear response between the potency and coagulation time of the standard and sample,with correlation coefficients r>0.99.As for the accuracy and repeatability,the recovery rates of various concentrations detected in the stock solution were 101.0%,100.0%and 112.0%,respectively,with RSD values of 2.6%,4.0%and 0.0%,respectively.The recovery rates of various concentrations in finished product testing were 104.0%,94.7%and 112.0%,respectively,with RSD values of 1.9%,2.4%and 0.0%,respectively.As for the intermediate precision,the RSD were 4.5%and 3.7%,respectively.After treated with sample diluent,the sample was tested at room temperature for 6 hours and still ex-hibited relatively stable biological activity.Conclusion This detection method is accurate,stable,easy to operate and highly automated,and is suitable for detecting the potency of recombinant human coagulation factorⅦa for Injection.

关 键 词：注射用重组人凝血因子Ⅶa 凝固法 效价检测 

分 类 号：R457.1[医药卫生—治疗学] R446.113[医药卫生—临床医学] R973.2