Platform development for high-throughput optimization of perfusion processes: Part I: Implementation of cell bleeds in microwell plates
Platform development for high-throughput optimization of perfusion processes: Part I: Implementation of cell bleeds in microwell plates
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DOI：
10.1002/bit.28682
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发表时间：
2024-02-23
影响因子：
3.8
通讯作者：
Micheletti，Martina
中科院分区：
文献类型：
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作者：
Dorn，Marie;Klottrup-Rees，Kerensa;Micheletti，Martina
持续加工以增加生物制药的产量和改善产品质量，同时减少制造足迹的承诺具有变革性。开发和优化灌流操作需要筛选各种参数，使用台式生物反应器时，这是昂贵和耗时的。缩小模型(SDMS)是高通量数据生成和条件筛选的最可行的选择。然而，需要新的模拟灌流的SDM，使实验能够并行运行。在这项研究中，提出了一种使用微孔板(MWP)在半灌流模式下运行的方法，并实施了细胞放血步骤。CHO细胞系在24孔截留分子量(VW= 1.2 mL)中培养，并在4个高细胞密度(Hcd)设定点生长。通过手动进行细胞放血，然后在离心后进行全培养液交换，获得准稳态条件。此外，两个HCD设定点被放大(VW= 30 毫升)，比较平方的六孔深井平板(DWP)和摇瓶(SF)。这项评估显示了系统(DWP与 + SF)和规模(MWP与DWP Sf)之间的可比性结果。结果表明，基于井板的方法适合于HCD和准稳态培养，为细胞克隆和培养基选择等工业相关挑战提供了强有力的解决方案。
The promise of continuous processing to increase yields and improve product quality of biopharmaceuticals while decreasing the manufacturing footprint is transformative. Developing and optimizing perfusion operations requires screening various parameters, which is expensive and time‐consuming when using benchtop bioreactors. Scale‐down models (SDMs) are the most feasible option for high‐throughput data generation and condition screening. However, new SDMs mimicking perfusion are required, enabling experiments to be run in parallel. In this study, a method using microwell plates (MWP) operating in semi‐perfusion mode with an implemented cell bleed step is presented. A CHO cell line was cultivated in a 24‐well MWP (Vw= 1.2 mL) and grown at four high cell density (HCD) setpoints. Quasi steady‐state condition was obtained by manually performing cell bleeds followed by a total medium exchange after centrifugation. Further, two HCD setpoints were scaled up (VW= 30 mL), comparing a squared six‐well deepwell plate (DWP) to shake flasks (SF). This evaluation showed comparable results between systems (DWP vs. SF) and scales (MWP vs. DWP + SF). The results show that the well‐plate‐based methods are suitable to perform HCD and quasi steady‐state cultivations providing a robust solution to industrially relevant challenges such as cell clone and media selection.