Dynamics of immature mAb glycoform secretion during CHO cell culture: An integrated modelling framework
Dynamics of immature mAb glycoform secretion during CHO cell culture: An integrated modelling framework
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DOI：
10.1002/biot.201400663
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发表时间：
2016-05-01
影响因子：
4.7
通讯作者：
Weilguny, Dietmar
中科院分区：
文献类型：
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作者：
del Val, Ioscani Jimenez;Fan, Yuzhou;Weilguny, Dietmar
鉴于结合在这些分子上的碳水化合物(多糖)的分布和组成决定了它们的治疗效果和免疫原性，确保治疗性单抗(MAbs)的糖基化相关质量已成为药物生物处理中的优先事项。然而，生物过程条件、细胞代谢和细胞内糖基化过程之间的相互作用仍有待充分了解。为了进一步了解这些相互作用，我们提出了一个新的集成建模平台，将mAb糖基化的动态变化与细胞分泌能力联系起来。比较了单抗特异性生产力(Qp)如何影响糖基化的两种不同的机制表示法。在第一种情况下，mAb的糖基化受分泌的货物通过高尔基体的线速度的调节。在第二种情况下，糖基化受到高尔基体体积变化的影响。在我们的建模框架内，这两种机制都准确地复制了实验观察到的mAb糖基化的动态变化。此外，还开发了一种基于优化的策略来估计将单抗糖型变异性降至最低所需的糖基化酶的浓度。我们的结果表明，相对于细胞分泌能力而言，糖基化机制的可用性可能在mAb糖基化过程中起着关键作用。在未来，这里提出的建模框架可能有助于选择和设计确保mAb糖基化一致的细胞系。
Ensuring consistent glycosylation‐associated quality of therapeutic monoclonal antibodies (mAbs) has become a priority in pharmaceutical bioprocessing given that the distribution and composition of the carbohydrates (glycans) bound to these molecules determines their therapeutic efficacy and immunogenicity. However, the interaction between bioprocess conditions, cellular metabolism and the intracellular process of glycosylation remains to be fully understood. To gain further insight into these interactions, we present a novel integrated modelling platform that links dynamic variations in mAb glycosylation with cellular secretory capacity. Two alternative mechanistic representations of how mAb specific productivity (qp) influences glycosylation are compared. In the first, mAb glycosylation is modulated by the linear velocity with which secretory cargo traverses the Golgi apparatus. In the second, glycosylation is influenced by variations in Golgi volume. Within our modelling framework, both mechanisms accurately reproduce experimentally‐observed dynamic changes in mAb glycosylation. In addition, an optimisation‐based strategy has been developed to estimate the concentration of glycosylation enzymes required to minimise mAb glycoform variability. Our results suggest that the availability of glycosylation machinery relative to cellular secretory capacity may play a crucial role in mAb glycosylation. In the future, the modelling framework presented here may aid in selecting and engineering cell lines that ensure consistent mAb glycosylatio.