Neunstoecklin Benjamin, Stettler Matthieu, Solacroup Thomas, Broly Hervé, Morbidelli Massimo, Soos Miroslav
Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.
Merck Serono S.A., Biotech Process Sciences, Corsier-sur-Vevey, Switzerland.
J Biotechnol. 2015 Jan 20;194:100-9. doi: 10.1016/j.jbiotec.2014.12.003. Epub 2014 Dec 18.
Application of quality by design (QbD) requires identification of the maximum operating range for parameters affecting the cell culture process. These include hydrodynamic stress, mass transfer or gradients in dissolved oxygen and pH. Since most of these are affected by the impeller design and speed, the main goal of this work was to identify a maximum operating range for hydrodynamic stress, where no variation of cell growth, productivity and product quality can be ensured. Two scale-down models were developed operating under laminar and turbulent condition, generating repetitive oscillating hydrodynamic stress with maximum stress values ranging from 0.4 to 420Pa, to compare the effect of the different flow regimes on the cells behavior. Two manufacturing cell lines (CHO and Sp2/0) used for the synthesis of therapeutic proteins were employed in this study. For both cell lines multiple process outputs were used to determine the threshold values of hydrodynamic stress, such as cell growth, morphology, metabolism and productivity. They were found to be different in between the cell lines with values equal to 32.4±4.4Pa and 25.2±2.4Pa for CHO and Sp2/0, respectively. Below the measured thresholds both cell lines do not show any appreciable effect of the hydrodynamic stress on any critical quality attribute, while above, cells responded negatively to the elevated stress. To confirm the applicability of the proposed method, the obtained results were compared with data generated from classical small-scale reactors with a working volume of 3L.
质量源于设计（QbD）的应用需要确定影响细胞培养过程的参数的最大操作范围。这些参数包括流体动力应力、传质或溶解氧和pH值的梯度。由于其中大多数参数受搅拌桨设计和转速的影响，因此本研究的主要目标是确定流体动力应力的最大操作范围，在此范围内可确保细胞生长、生产力和产品质量无变化。开发了两种在层流和湍流条件下运行的缩小模型，产生最大应力值范围为0.4至420Pa的重复性振荡流体动力应力，以比较不同流动状态对细胞行为的影响。本研究采用了两种用于合成治疗性蛋白质的生产细胞系（CHO和Sp2/0）。对于这两种细胞系，使用多个过程输出参数来确定流体动力应力的阈值，如细胞生长、形态、代谢和生产力。发现这两种细胞系的阈值不同，CHO细胞系和Sp2/0细胞系的值分别为32.4±4.4Pa和25.2±2.4Pa。在测量的阈值以下，两种细胞系均未显示流体动力应力对任何关键质量属性有明显影响，而在阈值以上，细胞对升高的应力有负面反应。为了证实所提出方法的适用性，将获得的结果与来自工作体积为3L的传统小规模反应器产生的数据进行了比较。
