Catabolic control for animal cell culture processes

哺乳动物细胞培养是制造许多重要的重组治疗性蛋白和疫苗的主要生产系统。尽管哺乳动物细胞技术取得了重大进展，但缺乏可靠的传感器，以及对细胞生理学和过程动力学的了解不足，阻碍了有效控制策略的发展。这些都是提高产量、保持产品质量和降低工艺可变性的关键和迫切需要。拟议的研究计划旨在开发基于模型的控制策略，使哺乳动物细胞培养过程能够稳健、高效和具有成本效益的开发和操作。更具体地说，该研究计划将研究1)使用定量代谢组学和13C-FLUX组学对细胞生理学进行动力学建模，2)关键生理变量的在线检测和估计3)能够保持细胞处于生理状态的喂养策略的设计和实施，以确保在培养的每个阶段都能保持高的生长和生产率。该方法将涉及基于广泛的生理特征的数学建模，使用质谱分析和同质异构体/代谢物平衡技术。该方法将首先在工业/临床相关的模型系统上开发。因此，这项赠款申请的目的与食品和药物管理局最近的“过程分析技术”倡议的要求直接一致。这一领域的进展对于增加临床新治疗剂的供应，同时缩短上市时间至关重要。通过产生基本和实际的好处，拟议的研究预计将为生物疗法的开发和制造带来重大贡献，生物疗法是一个具有相当重要经济意义的领域，直接影响加拿大人的健康和福利。

Mammalian cell cultures are the dominant production system for the manufacturing of many important recombinant therapeutic proteins and vaccines. Despite significant advances in mammalian cell technologies, the lack of reliable sensors and the insufficient understanding of cell physiology and process dynamics have hampered the development of effective control strategies. These are vital and critically needed for enhancing yield, maintaining product quality and reducing process variability. The proposed research program aims at developing model-based control strategies enabling the robust, efficient and cost-effective development and operation of mammalian cell culture processes. More specifically, the research program will investigate 1) the kinetic modeling of cellular physiology using quantitative metabolomics and 13C-fluxomics 2) the on-line sensing and estimation of key physiological variables 3) the design and implementation of feeding strategies capable of maintaining the cells at a physiological state that ensures high growth and production rates at each stage of culture. The methodology will involve mathematical modeling based on extensive physiological characterization using mass spectrometry analysis and isotopomer/metabolite balancing techniques. The approach will be developed initially on industrially/clinically relevant model systems. The aims of this grant application are thus in direct line with the requirements of the recent "Process Analytical Technology" initiative by the Food and Drug Administration. Advancements in this area are crucial to increase the supply of new therapeutic agents to the clinic, while reducing the time-to-market. By producing both fundamental and practical benefits, the proposed research is expected to bring a significant contribution to the development and manufacturing of biotherapeutics, an area of considerable economic importance that directly impact the health and welfare of Canadians.