抗原亲和色谱用于功能分离和有效表征关键单克隆抗体变体的功能分离和有效表征
Julia Baumeister1, Miriam Erhardt1, Johannes G Wilhelm2
1Global Development CMC Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany; Institute of Pharmaceutical Biotechnology, Ulm University, Ulm, Germany.
Journal of chromatography. A
|February 6, 2026
概括
使用抗原亲和色谱的新工作流将基于结合功能的单克隆抗体 (mAb) 变体进行分离. 这种方法可以识别出影响抗体有效性的关键修改,如异构化和顺丁胺的形成.
科学领域:
- 生物制药的发展.
- 分析化学是一种分析化学.
- 蛋白质化学 蛋白质化学
背景情况:
- 单克隆抗体 (mAbs) 经历翻译后的修改,影响产品质量.
- 补充性决定区域 (CDR) 的修改对抗原结合产生了重大影响,是关键的质量属性.
研究的目的:
- 开发一个模块化工作流程,以基于抗原亲和力的 mAb 变体的功能分离.
- 为了确定影响mAb结合的关键修改,并描述它们的影响.
主要方法:
- 用线性pH梯度 (pH7.4-2.5) 的分析抗原亲和色谱进行变异分离.
- 使用表面等离子共振 (SPR),SEC,IEC,CGE和映射对分离变体的表征.
- 工作流程的优化和应用到trastuzumab/Her2和两个额外的mAbs.
主要成果:
- 工作流成功地分离了绑定缺陷的mAb变体.
- 鉴定出HC-D102异构和LCN30糖胺 (Asu) 的形成是减少结合的关键修改.
- 该方法在不同的mAbs中表现出适应性,解决了高达66%的结合活性损失的变体.
结论:
- 开发的工作流允许有针对性地识别影响抗原结合的关键mAb修饰.
- 这种方法支持mAb开发中的早期风险缓解和控制策略.
- 实现了mAb变体的全面表征,确保了产品的质量和有效性.
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