Perfusion technology has been identified as a process improvement capable of eliminating some of the constraints in cell culture and allows for high cell densities and viabilities. However, when implementing this N‐1 seed perfusion platform in large‐scale manufacturing, unexpected cell damage was observed as early as Day 1. Given that the shear rate within recirculation hollow fibers was normalized and aligned correctly across bench, pilot, and manufacture scale, the primary mitigation was placed on the rotary lobe pump. Lowering the pump rate in manufacture scale successfully alleviated the cell damage. To understand the source of cell damage within the pump, a small‐scale rotary lobe‐pump robustness model was developed. Testing different pump flow rates and back pressures, it was concluded that high back pressure can cause cell damage. The back pressure within the system can cause back flow and high shear within small clearances inside the pump, which lead to the primary cell damage observed at a large scale. This shear level can be significantly higher than the shear in the hollow fiber. This pump robustness model can be utilized to aid the perfusion skid design, including pump operation efficiency and cell shear sensitivity. Methods to reduce the back pressure and cell shearing were determined to better predict manufacturing performance in the future.

中文翻译：

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开发用于检查大规模 N-1 种子灌注过程中细胞损伤的小型旋转叶泵细胞培养模型。

灌注技术已被确定为一种工艺改进，能够消除细胞培养中的一些限制，并允许高细胞密度和活力。然而，在大规模制造中实施这种 N-1 种子灌注平台时，早在第 1 天就观察到了意外的细胞损伤。 鉴于再循环中空纤维内的剪切速率在试验台、中试和制造规模上已标准化并正确对齐，主要缓解措施被放置在旋转凸轮泵上。降低制造规模的泵速成功地减轻了细胞损伤。为了了解泵内细胞损伤的来源，开发了一种小型旋转凸轮泵稳健性模型。测试不同的泵流量和背压，得出的结论是高背压会导致电池损坏。系统内的背压会导致泵内小间隙内的回流和高剪切，从而导致大规模观察到的原代细胞损伤。该剪切水平可以显着高于中空纤维中的剪切。该泵稳健性模型可用于辅助灌注橇设计，包括泵运行效率和细胞剪切敏感性。确定降低背压和电池剪切的方法以更好地预测未来的制造性能。包括泵运行效率和细胞剪切敏感性。确定降低背压和电池剪切的方法以更好地预测未来的制造性能。包括泵运行效率和细胞剪切敏感性。确定降低背压和电池剪切的方法以更好地预测未来的制造性能。