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CHO-S Antibody Titers 1 Gram/Liter Using Flow Electroporation-Mediated Transient Gene Expression followed by Rapid Migration to High-Yield Stable Cell Lines

机译:CHO-S抗体滴度 1克/升采用流电穿孔介导的瞬时基因表达然后快速迁移至高产稳定细胞系

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摘要

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.
机译:近年来,研究人员已将瞬态基因表达(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步,但大多数基于CHO的TGE系统产生的抗体滴度不足,无法在生物治疗开发流程中广泛使用。使用MaxCyte STX可扩展转染系统的流电穿孔是基于CHO的TGE的高效,可扩展手段,用于克级生产抗体,而无需专门的表达载体或基因工程的CHO细胞系。 CHO细胞流电穿孔可以轻松地从毫克放大到几克,而无需重新优化协议,同时保持转染性能和抗体生产率。本文提供的数据证明了使用MaxCyte STX的基于CHO的TGE的可重复性,可扩展性和抗体生产能力。数据显示转染后参数的优化,例如细胞密度,培养基组成和补料策略,可在单次CHO-S细胞转染后2周内导致分泌的抗体滴度> 1 g / L并产生数克抗体。另外,提供了数据来证明可分级电穿孔在快速产生高产率稳定CHO细胞系中的应用,以弥合早期和晚期抗体开发活性之间的差距。

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