英夫利西单抗糖化方法开发验证及其在抗体药物中的普适性  

Development and Validation of a Glycation Analysis Method for Infliximab and Its Universal Application Across Antibody Drugs

作　　者：徐梦娇 刘涛 徐进 郭清城 付蓉蓉 赵香 王福贵 宋兰坤 冀芦沙 钱卫珠 侯盛 李军 张大鹏 郭怀祖 XU Mengjiao;LIU Tao;XU Jin;GUO Qingcheng;FU Rongrong;ZHAO Xiang;WANG Fugui;SONG Lankun;JI Lusha;QIAN Weizhu;HOU Sheng;LI Jun;ZHANG Dapeng;GUO Huaizu;无(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;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]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155-168,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：糖化修饰作为一种非酶促糖修饰,通过还原性糖与蛋白氨基酸残基共价结合,普遍发生于抗体药物生产储存等过程,显著影响产品的质量和功能。硼酸亲和色谱(BAC)法虽然是检测抗体类药物糖化修饰的主要技术,但其在测定英夫利西单抗糖化水平时仍面临准确性挑战,在一定程度上限制了其在质量控制上的应用。本研究深入探讨了影响该单抗BAC方法准确性的因素,明确了该单抗BAC方法糖化检测不准确的原因,并针对性地对流动相的组成和浓度进行了优化,成功开发出一种改进的BAC方法,能够更精准地测定英夫利西单抗的糖化水平。该方法的准确性通过基于质谱的快速多属性方法确认,并通过了严格的方法学验证。验证结果表明该方法具有良好的准确性、精密度、耐用性等。此外,通过对不同类型抗体药物进行测试,证实了该方法在常规抗体药物糖化检测中的广泛适用性。本研究不仅解决了英夫利西单抗糖化检测的关键技术难题,还确认了该方法的普适性,为抗体药物的质量控制提供了一种新的快速、准确的技术方案。这些研究结果有助于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能力。Glycation,a pivotal non-enzymatic modification process characterized by the covalent attachment of reducing sugars to amino acid residues in proteins,is prevalent during the manufacturing and storage phases of antibody therapeutics.This process can critically impact drug quality and efficacy.Although boronate affinity chromatography(BAC)is a widely adopted method to assess glycation modifications in antibody drugs,its use in precisely quantifying infliximab glycation levels has encountered significant accuracy issues,impeding its effectiveness in quality control.Addressing this,our research dissected the elements influencing BAC precision for infliximab,pinpointing specific inaccuracies in glycation detection.Targeted optimization of the mobile phase constituents and concentrations culminated in the establishment of a refined BAC technique.This novel method allows for a more exact quantification of glycation levels in infliximab,with its reliability qualified by a mass spectrometry-based rapid multi-attribute approach,followed by rigorous methodological validation.Our results underscore the method’s exceptional accuracy,precision,and resilience.Extensive testing on a variety of antibody drugs further established the method’s versatile suitability for standard glycation assessment.This research not only overcomes seminal technical hurdles in infliximab glycation analysis but also affirms the method’s universal applicability.It introduces an expedited,precise strategy for antibody drug quality management,promising to expedite enhancements in technological frameworks and intellectual paradigms,thus fortifying product development and quality oversight capabilities.

关 键 词：英夫利西单抗 糖化 硼酸亲和色谱 抗体药物 质量控制 普适性 

分 类 号：R966[医药卫生—药理学]