The goal of cell culture process intensification is to increase volumetric productivity, generally by increasing viable cell density (VCD), cell specific productivity or production bioreactor utilization in manufacturing. In our previous study, process intensification in fed-batch production with higher titer or shorter duration was demonstrated by increasing the inoculation seeding density (SD) from ~ 0.6 (Process A) to 3–6 × 10⁶ cells/mL (Process B) in combination with media enrichment. In this study, we further increased SD to 10–20 × 10⁶ cells/mL (Process C) using perfusion N-1 seed cultures, which increased titers already at industrially relevant levels by 100% in 10–14 day bioreactor durations for four different mAb-expressing CHO cell lines. Redesigned basal and feed media were critical for maintaining higher VCD and cell specific productivity during the entire production duration, while medium enrichment, feeding strategies and temperature shift optimization to accommodate high VCDs were also important. The intensified Process C was successfully scaled up in 500-L bioreactors for 3 of the 4 mAbs, and quality attributes were similar to the corresponding Process A or Process B at 1000-L scale. The fed-batch process intensification strategies developed in this study could be applied for manufacturing of other mAbs using CHO and other host cells.

中文翻译：

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使用N-1灌注种子开发滴度加倍的强化补料分批生产平台，用于细胞培养

加强细胞培养过程的目的是通常通过提高活细胞密度（VCD），细胞比生产率或生产中生物反应器的利用率来提高容积生产率。在我们之前的研究中，通过将接种接种密度（SD）从〜0.6（过程A）提高到3–6×10 ⁶细胞/ mL（过程B），可以证明滴度更高或持续时间较短的分批生产中的过程强化。与媒体丰富化相结合。在这项研究中，我们将SD进一步提高到10–20×10 ⁶ 使用灌注的N-1种子培养物培养的细胞/ mL（过程C），对于四种不同的表达mAb的CHO细胞系，在10-14天的生物反应器持续时间内，其滴度已经在工业相关水平上提高了100％。重新设计的基础培养基和进料培养基对于在整个生产期间保持较高的VCD和细胞比生产力至关重要，同时培养基的富集，进料策略和温度变化的优化以适应高VCD也很重要。增强后的过程C已成功在500升生物反应器中按比例放大了4个mAb中的3个，并且质量属性类似于相应的过程A或过程B在1000升规模。本研究中开发的补料分批工艺强化策略可用于使用CHO和其他宿主细胞生产其他mAb。