摘要
目的:采用成像毛细管等电聚焦电泳(imaging capillary electrofocusing electrophoresis,iCIEF)分析单抗的电荷异质性,组织国内多家质控实验室对该方法进行验证;制备用于iCIEF法的系统适用性对照品。方法:邀请19个质控实验室,应用3种品牌(4种型号)的iCIEF设备进行方法学的预验证,确定该方法的关键试剂、仪器参数设置、数据分析要点等;依据ICH_Q2_R1指导原则、2015年版《中华人民共和国药典》(简称《中国药典》)通则9101制定方法学验证方案,邀请11个质控实验室参加验证;制备基于iCIEF法的系统适用性对照品并利用SEC-HPLC、IEC-HPLC、非还原CE-SDS进行稳定性评价。结果:预验证结果显示:等电点标记物(pI Marker)为关键试剂;不同品牌、型号的iCIEF检测设备参数设置无需统一;数据处理可采用自动积分、手动积分相结合的形式。验证结果:完成了该方法的特异性、准确度、精密度、线性及其范围、耐用性、定量下限的评价;制备了iCIEF法的系统适用性对照品,并进行了加速及长期稳定性评价,制定了相应的合格标准。结论:首次组织了iCIEF的方法学验证,为《中国药典》标准提高提供方法学验证依据,为该方法制备了系统适用性对照品。
Objective:To use imaging capillary electrofocusing electrophoresis(iCIEF) for charge heterogeneity analysis of monoclonal antibody products,to organize multiple quality control laboratories in China to carry out validation of the method and prepare system suitability control materials for iCIEF. Methods:Three brands(four models) of iCIEF equipments were used to carry out the pre-validation of methodology of iCIEF,and to determine the key reagents,instrument parameters,data analysis key points of the method in 19 quality control laboratories. According to ICH_Q2_R1 guideline and general principles 9101 of the 2015 edition of the Pharmacopoeia of the People’s Republic of China(China Pharmacopoeia),a protocol of validation for methodologies was formulated. Eleven quality control laboratories were invited to participate in the validation;the system suitability control materials based on iCIEF method were prepared and their stability was evaluated by SEC-HPLC,IEC-HPLC and non-reduced CE-SDS. Results:The pre-validation results showed that the isoelectric point marker(pI Marker) was the key reagent,it was not necessary to unify the parameters of iCIEF detection equipment of different brands and models,and data processing could adopt the combination of automatic and manual integration. The methodological validation results:the specificity,accuracy,precision,linearity and range,durability and quantitative limits of the method were evaluated,the system suitability control materials for iCIEF method were successfully prepared,its acceleration and long-term stability were evaluated,and the corresponding qualification standards were formuated. Conclusion:The validation of iCIEF methodology was organized for the first time,to provide a methodological validation basis for the improvement of China Pharmacopoeia standards,and a system suitability control material was prepared for the method.
作者
武刚
于传飞
王文波
李萌
王兰
WU Gang;YU Chuan-fei;WANG Wen-bo;LI Meng;WANG Lan(Division of Monoclonal Antibody,National Institutes for Food and Drug Control,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,Beijing 102629,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2019年第1期62-69,共8页
Chinese Journal of Pharmaceutical Analysis
基金
2017中国药典标准提高课题-63(单抗电荷变异体测定法-成像毛细管等电聚焦电泳法的建立)
关键词
单克隆抗体
电荷异质性
系统适用性对照品
成像毛细管等
电聚焦电泳(iCIEF)
方法学验证
monoclonal antibody
charge heterogeneity
system suitability control materials
imaging capillary electrofocusing electrophoresis (iCIEF )
method validation
