多糖基化位点抗体药物质控中基于单四级杆的多属性分析方法开发与验证  

Development and Validation of a Quadrupole Dalton-Based Multi-Attribute Method for the Quality Control of an Antibody Drug with Multiple Glycosylation Sites

作　　者：王福贵 刘涛 李锦豪 徐梦娇 付蓉蓉 赵香 聂爱英 宋兰坤 陈熙 冀芦沙 钱卫珠 侯盛 李军 徐忠升 范文强 张大鹏 徐进 郭清城 WANG Fugui;LIU Tao;LI Jinhao;XU Mengjiao;FU Rongrong;ZHAO Xiang;NIE Aiying;SONG Lankun;CHEN Xi;JI Lusha;QIAN Weizhu;HOU Sheng;LI Jun;XU Zhongsheng;FAN Wenqiang;ZHANG Dapeng;XU Jin;GUO Qingcheng(State Key Laboratory of Macromolecular Drugs and Large-scale Manufacturing,School of Pharmaceutical Sciences,Liaocheng University,Liaocheng 252059,China;NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China;State Key Laboratory of Macromolecular Drugs and Large-scale Manufacturing,School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China;State Key Laboratory of Macromolecular Drugs and Large-scale Manufacturing,Shanghai Zhangjiang Biotechnology Co.,Ltd.,Shanghai 201203,China;Department of Oncology,Huashan Hospital,Fudan University,Shanghai 200040,China;Taizhou Mabtech Pharmaceuticals Co.,Ltd.,Taizhou 225316,China;Waters Corporation,Shanghai 200126,China;Agilent Technologies(China)Co.,Ltd.,Shanghai 200131,China)

机构地区：[1]聊城大学药学院大分子药物与规模化制备全国重点实验室,山东聊城252059 [2]国家药品监督管理局治疗类单抗质量控制重点实验室,上海201203 [3]温州医科大学药学院大分子药物与规模化制备全国重点实验室,浙江温州325035 [4]张江生物技术有限公司大分子药物与规模化制备全国重点实验室,上海201203 [5]复旦大学华山医院肿瘤科,上海200040 [6]泰州迈博太科药业有限公司,江苏泰州225316 [7]沃特世科技(上海)有限公司,上海200126 [8]安捷伦科技(中国)有限公司,上海200131

出　　处：《聊城大学学报（自然科学版）》2024年第5期169-182,共14页Journal of Liaocheng University:Natural Science Edition

基　　金：国家重点研发计划项目(2021YFC2501700);国家自然科学基金项目(22304067);山东省自然科学基金项目(ZR2021LSW001,ZR2021MC017,ZR2021MB097,ZR2020KH019,ZR20230B257,ZR2023QB196)资助。

摘　　要：抗体药物已经成为生物制品的重要组成部分,此类产品具有复杂的质量属性,尤其是具有多糖基化位点的抗体药物,其质量控制面临更高挑战。虽然基于高分辨质谱的多属性分析方法(Multi-attribute method,MAM)已在抗体药物的质控中获得应用,但针对多糖基化位点抗体药物的快速且简便的检测技术仍较为少见。开发一种基于单四级杆的多属性分析方法,对于提升这类药物的质控效率具有重要意义。本研究以双糖基化位点的治疗类单抗CMAB009为模型。通过基于高分辨质谱的MAM对其进行了深入的预表征分析,成功获取了其主要质量属性。随后将预表征信息用于基于单四级杆的MAM开发。该方法在单次分析中有效监测了多种产品质量属性,其准确性通过基于高分辨质谱的方法进行了确认。随后我们对该方法进行了严格的方法学验证。验证结果显示,该方法具有良好的专属性、线性、准确度以及精密度等。基于单四级杆的MAM不仅有效地解决了多糖基化位点单抗药物的多属性检测难题,还展示了一定的方法普适性,为不同糖基化位点抗体药物的质量控制提供了一种新的、简便的技术方案。本研究结果预计将推动相关技术平台的改进和理念的更新,从而增强产品开发和质控的整体能力。Antibody drugs are pivotal in biopharmaceuticals,featuring complex quality attributes that complicate quality control,especially those with multiple glycosylation sites.Although high-resolution mass spectrometry-based multi-attribute methods(MAM)are established for quality control,rapid and straightforward testing methods for such antibodies are limited.This study introduces a Quadrupole Dalton-based MAM,enhancing quality control efficiency significantly.We used the therapeutic monoclonal antibody CMAB009,with dual glycosylation sites,as a model.Extensive pre-characterization with high-resolution MAM captured its primary quality attributes effectively.This data informed the development of a Quadrupole Dalton-based MAM that successfully monitored multiple quality attributes in a single analysis.Its accuracy,verified against high-resolution mass spectrometry,and thorough methodological validation,confirmed its high specificity,linearity,accuracy,and precision.This Quadrupole Dalton-based MAM not only addresses the challenges in multi-attribute testing of monoclonal antibodies with multiple glycosylation sites but also offers a new,simplified approach to quality control for various antibody drugs.This research is poised to enhance technology platforms and inspire conceptual innovations,thus boosting overall product development and quality control capabilities.

关 键 词：抗体药物 多属性分析方法 质量控制 CMAB009 单四级杆 

分 类 号：R917[医药卫生—药物分析学]