CE方法用于mAbs和复杂格式生物治疗药物的电荷变异分析
Maximilian Meudt1,2, Martin Pannek1,3, Nina Glogowski1
1Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.
Electrophoresis
|January 17, 2024
概括
我们开发了两种毛细电泳方法,用于分析治疗单克隆抗体 (mAbs) 和复杂格式的电荷异质性. 这些优化的方法提高了分辨率,并为mAb分析提供了强大的框架.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 生物制药学分析
背景情况:
- 单克隆抗体 (mAbs) 的电荷异质性影响治疗疗效和安全性.
- 分析像 bispecifics 这样的复杂的 mAb 格式带来了独特的分析挑战.
- 现有的方法可能缺乏解决方案或范围,以进行全面的负荷异质性评估.
研究的目的:
- 开发和优化基于毛细管电泳 (CE) 的方法,用于分析治疗 mAbs 和复杂格式的电荷异质性.
- 为了提高毛细管区电泳 (CZE) 和成像毛细管异电聚焦 (iCIEF) 的分辨率和分析范围.
主要方法:
- 开发一种新的缓冲系统,并优化CZE的背景电解质 (BGE),包括动态涂料剂和聚合物添加剂.
- 优化iCIEF使用特定的合物组合用于线性pH梯度,并研究非变性稳定剂.
- 在CZE中增加BGE pH,以改善高PI和复杂的mAb变体的分离.
主要成果:
- 优化的CZE方法证明了对高PI和复杂格式mAbs.的增强分辨率.
- 优化的iCIEF方法提供了适合广泛的mAb分析的线性pH梯度.
- 非洗剂的硫贝195在ICIEF中被确定为退化稳定剂的有效替代品.
结论:
- 开发的 CZE 和 iCIEF 方法为治疗 mAbs 的综合性电荷异质性分析提供了强大的框架.
- 这些优化技术改善了对生物制药开发的关键质量属性的表征.
- 该研究为各种mAb格式的常规电荷异质性评估提供了基础.
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