Platform development for high-throughput optimization of perfusion processes-Part II: Variation of perfusion rate strategies in microwell plates
Platform development for high-throughput optimization of perfusion processes-Part II: Variation of perfusion rate strategies in microwell plates
复制标题
灌注过程高通量优化的平台开发第二部分：微孔板灌注速率策略的变化
DOI：
10.1002/bit.28685
复制
发表时间：
2024
影响因子：
3.8
通讯作者：
Dorn M
中科院分区：
文献类型：
--
作者：
Dorn M
生物制药行业正在用灌注工艺取代补料分批工艺，以利用由于高细胞密度（HCD）操作和提高生产率而降低的资本和运营成本。HCD通过细胞保留和连续培养基交换来实现，这通常基于细胞特异性灌注率（CSPR）。为了获得成本生产工艺，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劳动密集型和耗时的实验，因为研究在实验室规模的灌注生物反应器中进行。微孔板（MWP）等小规模模型为在半灌注模拟中并行筛选多种灌注速率提供了一种选择。本研究研究了应用于在半灌注中运行的MWP平台的两种灌注速率策略。基于CSPR的灌注速率策略旨在整个培养过程中保持多个CSPR值，并与灌注速率为1 RV d−1的培养进行比较。研究细胞性能的双重目的是（i）在常规和HCD下进行细胞分离时实现HCD，以及（ii）在应用额外的手动细胞分离时保持HCD。采用两种灌注速率策略，活细胞浓度达到50 × 106cells mL− 1，并且获得了关键代谢物和抗体产物滴度的可比结果。此外，细胞流出和基于CSPR的培养基交换的组合应用成功地显示出分别在生长、代谢物和生产率方面的相似结果，同时将HCD培养的培养基消耗减少高达50%。
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 × 106cells mL−1were 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.