单抗药物分子大小变异体分析超速离心法的建立  

Establishment of an analytical ultracentrifugation method for analyzing size variants ofmonoclonal antibodies

作　　者：俞小娟 褚文丹[2] 余启昆 王飞宇 崔永霏 常卿 叶苗 霍德华 李文奇[2] 于传飞[1] 王兰[1] YU Xiaojuan;CHU Wendan;YU Qikun;WANG Feiyu;CUI Yongfei;CHANG Qing;YE Miao;HUO Dehua;LI Wenqi;YU Chuanfei;WANG Lan(Division of Monoclonal Antibodies,National Institutes for Food and Drug Control,State Key Laboratory of Drug Regulatory Science,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;Technology Center for Protein Sciences,Tsinghua University,Beijing Frontier Research Center for Biological Structure,Tsinghua University,Beijing 100084,China;Beckman Coulter Life Sciences,Shanghai 200131,China;School of Pharmaceutical Engineering,Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]中国食品药品检定研究院单克隆抗体产品室/药品监管科学全国重点实验室/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/国家药品监督管理局生物制品质量研究与评价重点实验室,北京102629 [2]清华大学蛋白质研究技术中心/清华大学北京生物结构前沿研究中心,北京100084 [3]贝克曼库尔特生命科学,上海200131 [4]沈阳药科大学制药工程学院,110016

出　　处：《中国医药生物技术》2025年第1期56-63,共8页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(21702007);国家重点研发计划“前沿生物技术”重点专项(2023YFC3404004)。

摘　　要：目的建立一种可用于测定单抗药物分子大小变异体的分析超速离心法。方法通过对转速、温度及数据分析参数等关键指标的优化,得到一种测定单抗药物分子大小变异体的方法,对该方法的专属性、重复性、精密度及线性进行方法学验证。结果优化得到的最佳实验条件:转速为50000 r/min,温度为20℃,分析参数“Resolution”为150,“s max”为20。方法学验证结果表明,6次重复性实验的RSD为0.91%,总体精密度RSD为0.78%,在0.2~0.8 mg/mL的浓度条件下线性良好。结论该分析超速离心法的专属性、重复性、精密度和线性良好,可用于单抗药物分子大小变异体的测定。Objective To establish an analytical ultracentrifugation method for measuring the size variants of monoclonal antibodies(mAb).Methods By optimizing key indicators such as rotational speed,temperature,and data analysis parameters,a method for measuring the size variants of mAbs was developed.The specificity,repeatability,precision,and linearity of this method were validated.Results The optimized experimental conditions were as follows:rotational speed at 50000 r/min,temperature at 20℃,analysis parameter resolution at 150,and s max at 20.Method validation results showed that the relative standard deviation(RSD)of six repeatability experiments was 0.91%,overall precision RSD was 0.78%,and the method demonstrated good linearity in the concentration range of 0.2-0.8 mg/mL.Conclusion The analytical ultracentrifugation method demonstrated good specificity,repeatability,precision,and linearity,making it suitable for determining the size variants of monoclonal antibodies.

关 键 词：单克隆抗体 纯度 分析超速离心 方法学验证 

分 类 号：R917[医药卫生—药物分析学]