Marschall Lukas, Gottimukkala Chitti Babu, Kayal Biswajit, Veeraraghavan Veerabhadra Madurai, Mandal Samir Kumar, Bandyopadhyay Suman, Herwig Christoph
TU Wien, Faculty of Technical Chemistry, Research Unit Biochemical Engineering, Gumpendorferstrasse 1a, 1060 Vienna, Austria.
Körber Pharma Austria GmbH, Mariahilfer Straße 88A/1/9, 1070 Vienna, Austria.
Bioengineering (Basel). 2023 Sep 30;10(10):1149. doi: 10.3390/bioengineering10101149.
Temperature downshifts are the gold standard when setting up control strategies for mammalian cell culture processes. These shifts are performed to prolong production phases and attain heightened levels of productivity. For the development of biosimilars, however, the bottleneck is in achieving a prespecified product quality. In a late-stage development project, we investigated the impact of temperature shifts and other process parameters with the aim of optimizing the glycosylation profile of a monoclonal antibody (mAb). We applied a design of experiments approach on a 3 L scale. The optimal glycosylation profile was achieved when performing a temperature upshift from 35.8 °C to 37 °C. Total afucosylated glycan (TAF) decreased by 1.2%, and galactosylated glycan species (GAL) increased by up to 4.5%. The optimized control strategy was then successfully taken to the manufacturing scale (1000 L). By testing two sets of set points at the manufacturing scale, we demonstrated that the statistical models predicting TAF and GAL trained with small-scale data are representative of the manufacturing scale. We hope this study encourages researchers to widen the screening ranges in process development and investigate whether temperature upshifts are also beneficial for other mAbs.
在为哺乳动物细胞培养过程制定控制策略时，温度下降是金标准。进行这些温度变化是为了延长生产阶段并实现更高的生产力水平。然而，对于生物类似药的开发而言，瓶颈在于实现预定的产品质量。在一个后期开发项目中，我们研究了温度变化及其他工艺参数的影响，旨在优化单克隆抗体（mAb）的糖基化谱。我们在3升规模上采用了实验设计方法。当从35.8°C升温至37°C时，实现了最佳糖基化谱。总去岩藻糖基化聚糖（TAF）降低了1.2%，半乳糖基化聚糖种类（GAL）增加了高达4.5%。然后，优化后的控制策略成功应用于生产规模（1000升）。通过在生产规模上测试两组设定点，我们证明了用小规模数据训练的预测TAF和GAL的统计模型能够代表生产规模。我们希望这项研究能鼓励研究人员在工艺开发中扩大筛选范围，并研究升温对其他单克隆抗体是否也有益。
