CHO-S antibody titers >1 gram/liter using flow electroporation-mediated transient gene expression followed by rapid migration to high-yield stable cell lines.
CHO-S antibody titers >1 gram/liter using flow electroporation-mediated transient gene expression followed by rapid migration to high-yield stable cell lines.
复制标题
CHO-S抗体滴度> 1克/升使用流动电穿孔介导的瞬时基因表达，然后快速迁移到高收益稳定的细胞系。
DOI：
10.1177/1087057114563494
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发表时间：
2015-04
影响因子：
--
通讯作者：
Peshwa M
中科院分区：
文献类型：
--
作者：
Steger K;Brady J;Wang W;Duskin M;Donato K;Peshwa M
近年来，研究人员已经转向瞬时基因表达（TGE）作为CHO稳定细胞系生成的替代方法，用于早期抗体开发。尽管在转染方法和培养优化方面取得了进展，但大多数基于CHO的TGE系统产生的抗体滴度不足以在生物制品开发管道中广泛使用。使用MaxCyte STX Scalable Transfection System的流式电穿孔是一种高效、可扩展的基于CHO的TGE方法，用于克级抗体生产，而无需专门的表达载体或基因工程CHO细胞系。CHO细胞流电穿孔很容易从毫克量扩展到数克量，而无需方案重新优化，同时保持转染性能和抗体生产率。在这篇文章中，数据显示，使用MaxCyte STX的CHO为基础的TGE的再现性，可扩展性和抗体生产能力。数据显示转染后参数如细胞密度、培养基组成和补料策略的优化，其导致分泌的抗体滴度> lg/L，并且在单次CHO-S细胞转染的2周内产生多克抗体。此外，还提供了数据，以证明可扩展电穿孔在快速生成高产稳定CHO细胞系中的应用，以弥合早期和晚期抗体开发活动之间的差距。
In recent years, researchers have turned to transient gene expression (TGE) as an alternative to CHO stable cell line generation for early-stage antibody development. Despite advances in transfection methods and culture optimization, the majority of CHO-based TGE systems produce insufficient antibody titers for extensive use within biotherapeutic development pipelines. Flow electroporation using the MaxCyte STX Scalable Transfection System is a highly efficient, scalable means of CHO-based TGE for gram-level production of antibodies without the need for specialized expression vectors or genetically engineered CHO cell lines. CHO cell flow electroporation is easily scaled from milligram to multigram quantities without protocol reoptimization while maintaining transfection performance and antibody productivity. In this article, data are presented that demonstrate the reproducibility, scalability, and antibody production capabilities of CHO-based TGE using the MaxCyte STX. Data show optimization of posttransfection parameters such as cell density, media composition, and feed strategy that result in secreted antibody titers >1 g/L and production of multiple grams of antibody within 2 weeks of a single CHO-S cell transfection. In addition, data are presented to demonstrate the application of scalable electroporation for the rapid generation of high-yield stable CHO cell lines to bridge the gap between early- and late-stage antibody development activities.