Catabolic control for animal cell culture processes

哺乳动物细胞培养物是制造许多重要的重组治疗性蛋白质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。这些对于提高产量、保持产品质量和减少工艺变异性至关重要。拟议的研究计划旨在开发基于模型的控制策略，从而实现哺乳动物细胞培养过程的稳健、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-通量组学进行细胞生理学动力学建模 2) 关键生理变量的在线传感和估计 3) 能够将细胞维持在生理状态的补料策略的设计和实施，从而确保每个培养阶段的高生长和生产率。该方法将涉及基于使用质谱分析和同位素异构体/代谢物平衡技术的广泛生理特征的数学建模。该方法最初将在工业/临床相关模型系统上开发。因此，这项拨款申请的目的直接符合美国食品和药物管理局最近发起的“过程分析技术”倡议的要求。这一领域的进步对于增加临床新治疗药物的供应、同时缩短上市时间至关重要。通过产生根本和实际效益，拟议的研究预计将为生物治疗药物的开发和制造做出重大贡献，这是一个具有相当大的经济重要性的领域，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.