摘要
目的根据ICH Q2(R^(2))分析方法验证指导原则的相关要求,对拟收录《中国药典》三部(2025版)的单克隆抗体(简称单抗)分子大小变异体分析方法分子排阻高效液相色谱(size-exclusion high-performance liquid chromatography,SEC-HPLC)法进行方法验证。方法首先制定SEC-HPLC分析方法验证方案、可接受标准以及分析线性和仪器线性样品,再对方法进行专属性、范围、准确度和精密度验证;最后对试验结果进行分析统计,得到单抗SEC-HPLC分析方法验证报告。结果该SEC-HPLC方法专属性较好,聚体的可报告范围为0.21%~8.33%,片段的可报告范围为0.06%~2.58%,单体的可报告范围≥89.09%;该方法的工作范围为5~300μg;聚体定量限为0.21%,片段定量限为0.06%;线性R^(2)>0.99;平均回收率在80%~110%之间;重复性和中间精密度RSD<8%。结论基于SECHPLC的单抗分子大小变异体分析方法专属性、范围、准确度和精密度均符合可接受标准。
Objective To verify the size-exclusion high-performance liquid chromatography(SEC-HPLC)method for molecular size variant analysis in monoclonal antibodies which is to be included in the Chinese Pharmacopoeia(VolumeⅢ,2025 edition),in accordance with the requirements of the ICH Q2(R^(2))Guideline for Analytical Method Validation.Methods Firstly,the validation plan,acceptable standards,and samples for analytical and instrument linearity were established.Then,the method was validated for specificity,range,accuracy and precision.Finally,the statistical analysis of test results was conducted to obtain the validation report of the SEC-HPLC method.Results The SEC-HPLC method exhibited a good specificity,with a reportable range of 0.21%to 8.33%for aggregates,0.06%to 2.58%for fragments,and≥89.09%for monomers.The working range of the method was 5-300μg,with limit of quantitation(LOQ)of 0.21%for aggregates and0.06%for fragments.The linear correlation coefficient(R^(2))exceeded 0.99.The mean recovery rates ranged between 80%and 110%.The RSDs for both repeatability and intermediate precision analyses were less than 8%.Conclusion The specificity,range,accuracy,and precision of the analytical method for molecular size variants in monoclonal antibodies based on SEC-HPLC all meet the acceptance criteria.
作者
杨雅岚
刘冰
崔永霏
李萌
武刚
于传飞
YANG Yalan;LIU Bing;CUI Yongfei;LI Meng;WU Gang;YU Chuanfei(Division of Monoclonal Antibodies,National Institutes for Food and Drug Control,NHC Key Laboratory of Research on Quality and Standardization of Biotech Products,NMPA Key Laboratory for Quality Research and Evaluation of Biological Products,Beijing 102629,China;不详)
出处
《中国生物制品学杂志》
2025年第11期1358-1365,共8页
Chinese Journal of Biologicals
