A dynamic mathematical model for monoclonal antibody N-linked glycosylation and nucleotide sugar donor transport within a maturing Golgi apparatus
A dynamic mathematical model for monoclonal antibody N-linked glycosylation and nucleotide sugar donor transport within a maturing Golgi apparatus
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DOI：
10.1002/btpr.688
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发表时间：
2011-11-01
影响因子：
2.9
通讯作者：
Kontoravdi, Cleo
中科院分区：
文献类型：
--
作者：
del Val, Ioscani Jimenez;Nagy, Judit M.;Kontoravdi, Cleo
单克隆抗体（mAb）是生物制药行业最重要的产品之一。它们的疗效取决于糖基化的翻译后过程，而糖基化过程受生产工艺条件的影响。在此，我们提出了一个动态的mAb糖基化的数学模型，考虑池成熟的高尔基体近似的活塞流反应器，并通过包括高尔基驻留蛋白（糖基化酶和转运蛋白[TP]）的回收。糖基化反应速率的表达是根据每种酶的动力学机制，从胞质溶胶到高尔基体腔的核苷酸糖供体[NSD]的运输建模，作为糖基化和细胞代谢之间的联系。开发了基于优化的方法来估计未知的酶和TP浓度分布参数。所得模型能够再现商业mAb的糖基化谱。它可以进一步再现FucT糖基化酶的基因沉默和胞质NSD消耗对mAb寡糖谱的影响。我们模型的所有新元素都基于生物学证据，并产生比以前报告更准确的结果。因此，我们认为，这些改进有助于更详细地表示N-连接的糖基化过程。总体结果显示了我们的模型对评估产生所需糖基化谱的细胞工程策略的潜力。此外，当与细胞代谢结合时，该模型可用于评估工艺条件对糖基化的影响，并有助于设计、控制和优化生物制药生产工艺。(C)2011年美国化学工程师学会生物技术。程序，2011
Monoclonal antibodies (mAbs) are one of the most important products of the biopharmaceutical industry. Their therapeutic efficacy depends on the post-translational process of glycosylation, which is influenced by manufacturing process conditions. Herein, we present a dynamic mathematical model for mAb glycosylation that considers cisternal maturation by approximating the Golgi apparatus to a plug flow reactor and by including recycling of Golgi-resident proteins (glycosylation enzymes and transport proteins [TPs]). The glycosylation reaction rate expressions were derived based on the reported kinetic mechanisms for each enzyme, and transport of nucleotide sugar donors [NSDs] from the cytosol to the Golgi lumen was modeled to serve as a link between glycosylation and cellular metabolism. Optimization-based methodologies were developed for estimating unknown enzyme and TP concentration profile parameters. The resulting model is capable of reproducing glycosylation profiles of commercial mAbs. It can further reproduce the effect gene silencing of the FucT glycosylation enzyme and cytosolic NSD depletion have on the mAb oligosaccharide profile. All novel elements of our model are based on biological evidence and generate more accurate results than previous reports. We therefore believe that the improvements contribute to a more detailed representation of the N-linked glycosylation process. The overall results show the potential of our model toward evaluating cell engineering strategies that yield desired glycosylation profiles. Additionally, when coupled to cellular metabolism, this model could be used to assess the effect of process conditions on glycosylation and aid in the design, control, and optimization of biopharmaceutical manufacturing processes. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011