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在蛋白质染色学过程中增强单克隆抗体的稳定性,使用2-甲基伊米达二酸

Ranjeet Desai¹, Rahul Jaiswal², Triveni Manchekar²

¹Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai 400019, India; Department of Biological Sciences and Biotechnology, Institute of Chemical Technology, Mumbai 400019, India.

Journal of chromatography. A

|August 18, 2024

概括

2-甲基伊米达二酸 (2-MIDHP) 在低pH的蛋白A净化过程中显著降低单克隆抗体 (mAb) 聚合. 这种添加剂增强了mAb的稳定性,而不会在生物处理过程中对回收或纯度产生负面影响.

科学领域:

生物制药制造业生物制药制造业
蛋白质化学蛋白质化学
过程开发过程开发

背景情况:

单克隆抗体 (mAb) 聚合是下游加工过程中的一个关键问题,可能会损害产品的质量和有效性.
低pH条件,通常用于蛋白质A染色体和病毒失活,可以加剧mAb的不稳定性和促进聚合.
在这些紧张条件下制定减轻聚合的策略对于稳健的生物制造至关重要.

研究的目的:

为了研究二甲基伊米达二酸盐 (2-MIDHP) 在低pH的蛋白A净化过程中对mAb聚合的影响.
为了评估2-MIDHP对mAb恢复,宿主细胞蛋白 (HCP) 清除和蛋白A漏的影响.
评估2-MIDHP对mAb热稳定性和结构完整性的影响.

主要方法:

使用大小排除高性能液态色谱 (SE-HPLC) 和动态光散射 (DLS) 来量化mAb聚合.
使用差分扫描热度计 (DSC) 来分析 mAb 的热稳定性.
循环二重化 (CD) 和光光谱学被用来探测mAb的结构变化.

主要成果:

2-MIDHP在pH值3.0时表现出度依赖的对mAb聚合的保护作用,随着度的增加,聚合率从3.8%降至0.56%.
没有观察到对mAb恢复,HCP清除或蛋白A漏的显著不良影响.
DSC,CD和光谱学表明,2-MIDHP增强了mAb的热稳定性,并保持了蛋白质结构,而不会导致变性.

结论:

2-MIDHP在低pH蛋白A净化过程中有效地减轻mAb聚合,可能是通过稳定抗体结构.
使用2-MIDHP是一个有前途的策略,可以提高mAb净化过程的稳定性和效率.
这些发现有助于提高治疗单克隆抗体的质量和产量.

关键词:

聚合是一种聚合.离子液体是一种离子液体.一个单克隆抗体的抗体.蛋白质A染色体的染色学净化净化净化净化

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