冻存对复溶后人粒细胞刺激因子国家标准品生物学活性的影响  

Effect of cryopreservation on biological activity of national standard of human granulocyte colony-stimulating factor after reconstitution

作　　者：朱留强 史新昌[1] 刘兰[1] 裴德宁[1] 于雷[1] 秦玺[1] 周勇[1] 王军志[1] ZHU Liuqiang;SHI Xinchang;LIU Lan;PEI Dening;YU Lei;QIN Xi;ZHOU Yong;WANG Junzhi(NHC Key Laboratory of Research on Quality and Standardization of Biotech Products,National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院、国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国生物制品学杂志》2023年第9期1093-1096,共4页Chinese Journal of Biologicals

基　　金：国家科技重大专项重大新药创制(2018ZX09101001-003);国家质量基础设施(National Quality Infrastructure,NQI)资助(2021YFF-0600804)。

摘　　要：目的探讨冻存对复溶后人粒细胞刺激因子(human granulocyte colony-stimulating factor,hG-CSF)国家标准品生物学活性的影响,为hG-CSF国家标准品的使用提供参考。方法依据《中国药典》三部(2020版)通则3525测定标准品的生物学活性;将复溶后的hG-CSF国家标准品分装后置于-80、-40、-20℃保存,分别于第1、2、3、5和6个月取样检测生物学活性,用临用前复溶的标准品定量-80℃存放不同时间的标准品,以-80℃存放的样品为100%活性对照品定量其余样品的相对生物学活性。结果热加速稳定试验拟合Eyrlng方程为:ln{k(t)}=6.75-3772.20/T+ln(T),R2=0.969,预测hG-CSF国家标准品于-80℃贮存时,生物学活性衰减5%约需93.4个月;-80℃冻存的复溶标准品生物学活性值下降约24%。结论hG-CSF国家标准品复溶分装后于-80℃可稳定保存半年以上。但冻融会引起活性值下降超过20%,因此不建议复溶后分装储存使用。Objective To investigate the effect of cryopreservation on the biological activity of the national standard of human granulocyte colony-stimulating factor(hG-CSF)after reconstitution,so as to provide a reference for the use of the national standard of hG-CSF.MethodsThe biological activity of the standard was determined according to the general rule 3525 of Chinese Pharmacopoeia(VolumeⅢ,2020 edition);The reconstituted hG-CSF national standard was aliquoted and stored at-80℃,-40℃and-20℃,and then sampled at 1,2,3,5 and 6 months to detect the biological activity.The standards reconstituted before use were used to quantify the standards stored at-80℃for different time durations,and the standards stored at-80℃were defined as the reference of 100%activity to quantify the relative biological activity of the other samples.ResultsThe Eyrlng equation fitted by the thermal acceleration stability experiment was:ln{k(t)}=6.75-3772.20/T+ln(T),R~2=0.969.The biological activity of hG-CSF national standard was predicted to decrease by 5%after about 93.4 months storage at-80℃;The biological activity of reconstituted standards frozen at-80℃decreased by about 24%.ConclusionThe aliquoted reconstituted hG-CSF national standard can be stored at-80℃stably for more than half a year.However,freezing and thawing will cause the activity value to drop by more than 20%,so it is not recommended to reuse after reconstitution.

关 键 词：复溶 人粒细胞刺激因子 国家标准品 生物学活性 

分 类 号：R917[医药卫生—药物分析学]