Development of a model-based optimization strategy for mab production

哺乳动物细胞培养是生产许多高价值生物药物的主要平台，包括酶、单抗、病毒疫苗、激素和免疫调节剂。本研究以中国仓鼠卵巢(CHO)细胞系为研究对象，采用流加分批培养的方法，建立了一种生产单抗的最佳生物处理方法。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。目标是找到在补料分批过程中实施的最佳补料配置和最佳操作条件，包括营养和温度配置。优化该系统的关键问题是防止或延缓被称为细胞凋亡的程序性细胞死亡。以前的研究只提出了该系统的部分模型。一些研究人员只对与细胞凋亡相关的信号转导通路进行了建模，而另一些研究人员则只对细胞外代谢的代谢通量进行了建模。目前的研究试图通过解释细胞外代谢产物通量的演变及其与细胞转导通路的特异性相互作用来阐明导致细胞凋亡的机制。这个用于优化的综合数学模型将通过对单抗生产过程中代谢物、细胞内和细胞外蛋白质水平以及活细胞、坏死细胞和凋亡细胞比例的实验监测来验证。预计通过调查和解决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，将产生一种系统的方法来获得生物制药生产的新的优化方案。对于目前昂贵的生物药品，这将为公众带来显著的成本节约。

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.