Development of a model-based optimization strategy for mab production

哺乳动物细胞培养是生产多种高价值生物药品的主要平台，包括酶、单克隆抗体（mab）、病毒疫苗、激素和免疫调节剂。本提案涉及开发一种新的基于优化的生物处理方法，用于中国仓鼠卵巢（CHO）细胞系使用饲料批量培养生产单抗。优化将基于该过程的详细模型，该模型将描述细胞外代谢物通量和细胞内的信号转导途径。目标将是找到一个最佳的饲料配置文件和最佳的操作条件，包括营养和温度配置文件，在饲料批次过程中实施。优化该系统的关键问题是预防或延迟程序性细胞死亡，即细胞凋亡。以前的研究只提出了该系统的部分模型。一些研究人员只模拟了与细胞凋亡相关的信号转导途径，而另一些研究人员只模拟了这一过程中的代谢细胞外通量。目前的研究旨在通过解释细胞外代谢物通量的进化及其与细胞凋亡转导途径的特定相互作用来阐明导致细胞凋亡的机制。该综合数学模型将用于优化，并将通过在Mab生产过程中对代谢物、细胞内和细胞外蛋白水平以及活细胞、坏死细胞和凋亡细胞比例的实验监测来验证。通过研究细胞外通量和与细胞凋亡相关的信号转导途径之间的相互作用，有望形成一种获得生物制药生产新最佳方案的系统方法。这将为公众节省目前昂贵的生物制药成本。

Mammalian cell culture comprises a major platform for producing a number of high-value biopharmaceuticals,including enzymes, monoclonal antibodies (Mabs), viral vaccines, hormones and immuno-regulators. Thisproposal deals with the development of a novel based optimal bioprocessing methodology for the production ofMab by the Chinese hamster ovary (CHO) cell line using fed batch culture. The optimization will be based on adetailed model of the process that will describe both the extracellular metabolite fluxes and the signaltransduction pathways in the cell. The objective will be to find an optimal feeding profile and optimal operatingconditions, including nutrient and temperature profile, to be implemented during the fed-batch process. Thekey issue for optimizing the system is the prevention or delay of programmed cell death referred to asapoptosis. Previous studies have proposed only partial models of the system. Some researchers only modelledthe signal transduction pathways related to apoptosis whereas others only modelled the metabolic extracellularfluxes during the process.The current research intends to elucidate the mechanisms leading to apoptosis by accounting for both theevolution of extracellular metabolite fluxes and their specific interaction with the cellular transductionpathways responsible for apoptosis. This comprehensive mathematical model, to be used for optimization, willbe validated by experimental monitoring of the levels of metabolites, intracellular and extracellular proteins aswell as the proportions of viable, necrotic and apoptotic cells during the Mab production process.It is expected that by investigating and addressing the interaction between the extracellular fluxes and thesignal transduction pathways realted to apoptosis, a systematic methodology for obtaining novel optimalprotocols for biopharmaceutical production will result. This will bring about a significant cost saving to thepublic regarding currently expensive biopharmaceuticals.