The biopharmaceutical industry is replacing fed‐batch with perfusion processes to take advantage of reduced capital and operational costs due to the operation at high cell densities (HCD) and improved productivities. HCDs are achieved by cell retention and continuous medium exchange, which is often based on the cell‐specific perfusion rate (CSPR). To obtain a cost‐productive process the perfusion rate must be determined for each process individually. However, determining optimal operating conditions remain labor‐intensive and time‐consuming experiments, as investigations are performed in lab‐scale perfusion bioreactors. Small‐scale models such as microwell plates (MWPs) provide an option for screening multiple perfusion rates in parallel in a semi‐perfusion mimic. This study investigated two perfusion rate strategies applied to the MWP platform operated in semi‐perfusion. The CSPR‐based perfusion rate strategy aimed to maintain multiple CSPR values throughout the cultivation and was compared to a cultivation with a perfusion rate of 1 RV d−1. The cellular performance was investigated with the dual aim (i) to achieve HCD, when inoculating at conventional and HCDs, and (ii) to maintain HCDs, when applying an additional manual cell bleed. With both perfusion rate strategies viable cell concentrations up to 50 × 106 cells mL−1 were achieved and comparable results for key metabolites and antibody product titers were obtained. Furthermore, the combined application of cell bleed and CSPR‐based medium exchange was successfully shown with similar results for growth, metabolites, and productivities, respectively, while reducing the medium consumption by up to 50% for HCD cultivations.

中文翻译：

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高通量优化灌注过程的平台开发——第二部分：微孔板灌注速率策略的变化

生物制药行业正在用灌注工艺取代分批补料，以利用高细胞密度 (HCD) 操作降低资本和运营成本并提高生产率。HCD 通过细胞保留和连续培养基更换来实现，这通常基于细胞特异性灌注率 (CSPR)。为了获得具有成本效益的工艺，必须单独确定每个工艺的灌注速率。然而，确定最佳操作条件仍然是一项劳动密集型且耗时的实验，因为研究是在实验室规模的灌注生物反应器中进行的。微孔板（MWP）等小规模模型提供了在半灌注模拟中并行筛选多个灌注速率的选择。本研究研究了应用于半灌注 MWP 平台的两种灌注速率策略。基于 CSPR 的灌注速率策略旨在在整个培养过程中维持多个 CSPR 值，并与灌注速率为 1 RV d 的培养进行比较−1。研究细胞性能的双重目的是 (i) 在常规和 HCD 接种时实现 HCD，以及 (ii) 在应用额外的手动细胞放血时维持 HCD。采用两种灌注速率策略时，活细胞浓度高达 50 × 106细胞毫升−1取得了关键代谢物和抗体产物滴度的可比结果。此外，细胞出血和基于 CSPR 的培养基交换的组合应用已成功地显示出在生长、代谢物和生产力方面分别具有相似的结果，同时将 HCD 培养的培养基消耗量减少了高达 50%。