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单克隆抗体充电变异的生物相似性和先进的结构特征使用毛细管区电泳和质谱学

Lola Alez-Martin¹, Pascal Houzé², Rania Joomun³

¹Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), Université de Strasbourg, CNRS UMR7140, CMC, Strasbourg, 67081, France; Université Paris Cité, CNRS, Inserm, Unité de Technologies Chimiques et Biologiques pour la Santé (UTCBS), 4 Avenue de l'observatoire, Paris, F-75006, France.

|June 25, 2025

概括

毛细管区域电泳 (CZE) 有效地分离由翻译后修饰引起的单克隆抗体 (mAb) 电荷变异. 这种方法有助于评估mAb的生物相似性,并识别各种结构变异.

科学领域:

生物化学生物化学
分析化学分析化学
生物制药产品生物制药产品

背景情况:

单克隆抗体 (mAbs) 由于其大小和翻译后修饰 (PTMs) 呈现结构复杂性,导致电荷变异.
毛细管区电泳 (CZE) 对mAb分析有价值,但其分离选择性需要进一步阐明.

研究的目的:

使用CZE-UV. 用于鉴定infliximab生物类似药中的充电变体.
调查充电变体的起源并评估mAb生物相似性.
开发一种分析策略,用于识别负责充电变体的PTM.

主要方法:

用紫外线检测 (CZE-UV) 进行毛细管区域电泳,用于分析因弗利西马布和潘布罗利祖马布的电荷变异.
用酶处理来归因充电变体的起源.
采集和丰富CZE分数与毛细体电泳-双重质谱学 (CE-MS/MS) 结合,用于离线表征.

主要成果:

CZE-UV成功地确定了因弗力西马布生物类似药的微妙变异,支持生物相似性评估.
对 pembrolizumab 的分析揭示了源自C端 lysine 或 N-glycans 的电荷变异.
CE-MS/MS 确定了 N-终端的高胺酸,阿斯帕拉金脱化 (与运动能力下降有关) 和苏胺的形成 (与运动能力增加有关).

结论:

通过分离电荷变体,CZE-UV提供了治疗mAb结构多样性的概述.
观察到的CZE-UV分离是多个PTM影响明显流动性的协同效应的结果.
开发的分析策略使mAb收费变体及其相关PTM的详细表征成为可能.

关键词:

毛细管区电泳是毛细管区的电泳.收费变种收费变种分数收集是分数的收集.质谱测量质量谱测量单克隆抗体是一种单克隆抗体.后翻译修改后的修改.

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