Abstract An alternative downstream process for the purification of a monoclonal antibody (mAb) based on aqueous two-phase extraction as clarification, capture and primary purification step was developed. For further purification unit operations which are commonly used for mAb platform processes were utilized. A diafiltration approach was used to combine virus inactivation and removal of phase forming components as well as low molecular weight impurities in one step followed by cation and anion exchange chromatography. Starting from cell containing cultivation broth, an overall mAb yield of 74 % was achieved within an application study. The process was optimized regarding mAb yield and the clearance of process related impurities like deoxyribonucleic acid as well as host cell proteins, which were removed to approximately 60 and 6000 ppm respectively. Critical product quality attributes, regarding glycosylation patterns, were also examined and remained unimpaired after the aqueous two-phase extraction. The alternative downstream process presented in this study offers great potential to improve mAb manufacturing.

中文翻译：

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基于水两相萃取纯化单克隆抗体的替代下游工艺

摘要 开发了一种基于水两相萃取作为澄清、捕获和初级纯化步骤的单克隆抗体 (mAb) 纯化的替代下游工艺。对于通常用于 mAb 平台过程的进一步纯化单元操作被利用。渗滤方法用于在一个步骤中结合病毒灭活和相形成组分以及低分子量杂质的去除，然后进行阳离子和阴离子交换层析。从含有细胞的培养液开始，在应用研究中实现了 74% 的总单克隆抗体产量。该工艺在 mAb 产量和工艺相关杂质（如脱氧核糖核酸以及宿主细胞蛋白）的清除方面进行了优化，分别去除到大约 60 和 6000 ppm。还检查了与糖基化模式相关的关键产品质量属性，并且在水相两相萃取后保持不变。本研究中介绍的替代下游工艺为改进 mAb 制造提供了巨大潜力。