The use of Process Analytical Technology tools coupled with chemometrics has been shown great potential for better understanding and control of mammalian cell cultivations through real‐time process monitoring. In‐line Raman spectroscopy was utilized to determine the glucose concentration of the complex bioreactor culture medium ensuring real‐time information for our process control system. This work demonstrates a simple and fast method to achieve a robust partial least squares calibration model under laboratory conditions in an early phase of the development utilizing shake flask and bioreactor cultures. Two types of dynamic feeding strategies were accomplished where the multi‐component feed medium additions were controlled manually and automatically based on the Raman monitored glucose concentration. The impact of these dynamic feedings was also investigated and compared to the traditional bolus feeding strategy on cellular metabolism, cell growth, productivity, and binding activity of the antibody product. Both manual and automated dynamic feeding strategies were successfully applied to maintain the glucose concentration within a narrower and lower concentration range. Thus, besides glucose, the glutamate was also limited at low level leading to reduced production of inhibitory metabolites, such as lactate and ammonia. Consequently, these feeding control strategies enabled to provide beneficial cultivation environment for the cells. In both experiments, higher cell growth and prolonged viable cell cultivation were achieved which in turn led to increased antibody product concentration compared to the reference bolus feeding cultivation.

中文翻译：

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在生产阿达木单抗的 CHO 细胞培养期间使用实时葡萄糖浓度监测系统的基于拉曼的动态补料策略。

过程分析技术工具与化学计量学的结合使用已显示出通过实时过程监测更好地理解和控制哺乳动物细胞培养的巨大潜力。在线拉曼光谱用于确定复杂生物反应器培养基的葡萄糖浓度，确保为我们的过程控制系统提供实时信息。这项工作展示了一种简单快速的方法，可在实验室条件下利用摇瓶和生物反应器培养物在开发的早期阶段实现稳健的偏最小二乘校准模型。实现了两种类型的动态进料策略，其中基于拉曼监测的葡萄糖浓度手动和自动控制多组分进料培养基的添加。还研究了这些动态补料的影响，并与传统的推注补料策略对抗体产品的细胞代谢、细胞生长、生产力和结合活性的影响进行了比较。手动和自动动态喂料策略均成功应用于将葡萄糖浓度保持在更窄和更低的浓度范围内。因此，除了葡萄糖，谷氨酸盐也被限制在低水平，导致抑制性代谢物的产生减少，如乳酸和氨。因此，这些饲养控制策略能够为细胞提供有益的培养环境。在这两个实验中，实现了更高的细胞生长和延长的活细胞培养，这反过来导致与参考团注补料培养相比抗体产物浓度增加。