Investigation of the interactions of critical scale-up parameters (pH, pO₂ and pCO₂) on CHO batch performance and critical quality attributes

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• 作者：Matthias Brunner ; Jens Fricke ; Paul Kroll…
• 关键词：CHO cell culture ; Process parameter ; Scale ; up ; Monoclonal antibody CQA ; Design of experiments
• 刊名：Bioprocess and Biosystems Engineering
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：40
• 期：2
• 页码：251-263
• 全文大小：1518KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Biotechnology; Industrial and Production Engineering; Environmental Engineering/Biotechnology; Industrial Chemistry/Chemical Engineering; Food Science;
• 出版者：Springer Berlin Heidelberg
• ISSN：1615-7605
• 卷排序：40

文摘

Understanding process parameter interactions and their effects on mammalian cell cultivations is an essential requirement for robust process scale-up. Furthermore, knowledge of the relationship between the process parameters and the product critical quality attributes (CQAs) is necessary to satisfy quality by design guidelines. So far, mainly the effect of single parameters on CQAs was investigated. Here, we present a comprehensive study to investigate the interactions of scale-up relevant parameters as pH, pO₂ and pCO₂ on CHO cell physiology, process performance and CQAs, which was based on design of experiments and extended product quality analytics. The study used a novel control strategy in which process parameters were decoupled from each other, and thus allowed their individual control at defined set points. Besides having identified the impact of single parameters on process performance and product quality, further significant interaction effects of process parameters on specific cell growth, specific productivity and amino acid metabolism could be derived using this method. Concerning single parameter effects, several monoclonal antibody (mAb) charge variants were affected by process pCO₂ and pH. N-glycosylation analysis showed positive correlations between mAb sialylation and high pH values as well as a relationship between high mannose variants and process pH. This study additionally revealed several interaction effects as process pH and pCO₂ interactions on mAb charge variants and N-glycosylation pattern. Hence, through our process control strategy and multivariate investigation, novel significant process parameter interactions and single effects were identified which have to be taken into account especially for process scale-up.