Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
Department of Pharmaceutical Science, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
Biotechnol Prog. 2023 Nov-Dec;39(6):e3368. doi: 10.1002/btpr.3368. Epub 2023 Jul 27.
A majority of the biotherapeutics industry today relies on the manufacturing of monoclonal antibodies from Chinese hamster ovary (CHO) cells, yet challenges remain with maintaining consistent product quality from high-producing cell lines. Previous studies report the impact of individual trace metal supplemental on CHO cells, and thus, the combinatorial effects of these metals could be leveraged to improve bioprocesses further. A three-level factorial experimental design was performed in fed-batch shake flasks to evaluate the impact of time wise addition of individual or combined trace metals (zinc and copper) on CHO cell culture performance. Correlations among each factor (experimental parameters) and response variables (changes in cell culture performance) were examined based on their significance and goodness of fit to a partial least square's regression model. The model indicated that zinc concentration and time of addition counter-influence peak viable cell density and antibody production. Meanwhile, early copper supplementation influenced late-stage ROS activity in a dose-dependent manner likely by alleviating cellular oxidative stress. Regression coefficients indicated that combined metal addition had less significant impact on titer and specific productivity compared to zinc addition alone, although titer increased the most under combined metal addition. Glycan analysis showed that combined metal addition reduced galactosylation to a greater extent than single metals when supplemented during the early growth phase. A validation experiment was performed to confirm the validity of the regression model by testing an optimized setpoint of metal supplement time and concentration to improve protein productivity.
当今，大多数生物制药行业依赖于中国仓鼠卵巢 (CHO) 细胞生产单克隆抗体，但在维持高产细胞系的一致产品质量方面仍存在挑战。先前的研究报告了个别痕量金属补充对 CHO 细胞的影响，因此，可以利用这些金属的组合效应进一步改进生物工艺。在分批补料摇瓶中进行了三水平析因实验设计，以评估逐个或组合添加痕量金属（锌和铜）对 CHO 细胞培养性能的影响。根据其对细胞培养性能变化（响应变量）的显著性和拟合度，检查了每个因素（实验参数）与响应变量之间的相关性。基于偏最小二乘回归模型。该模型表明，锌浓度和添加时间对峰值活细胞密度和抗体产量有反影响。同时，早期铜补充以剂量依赖的方式影响晚期 ROS 活性，可能通过减轻细胞氧化应激。回归系数表明，与单独添加锌相比，组合金属添加对效价和比生产率的影响较小，尽管在组合金属添加下效价增加最多。聚糖分析表明，与单独添加金属相比，在早期生长阶段补充组合金属会更大程度地降低半乳糖基化程度。进行了验证实验，通过测试金属补充时间和浓度的优化设定点来提高蛋白质产率，以验证回归模型的有效性。
