Characterization, modeling and environmental manipulation of mammalian cell culture systems for optimal protein 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。生物制药工业的未来将依赖于这些哺乳动物细胞培养生产系统的发展和增强。这类生物制剂的产量相对较小，因为所需的培养条件高度专业化，而且由于培养条件的轻微变化，培养物很容易降低生产率或导致显著的细胞死亡。早期的生物化学工程师专注于生产的非生物方面，如反应器设计，而最近的工程发展已经认识到，提高生产的最大潜力在于动物细胞本身。在本提案中，生物工艺开发的一个重要且经常被忽视的方面，即生物工艺优化被解决。这一建议涉及一种新的优化策略的发展，为生产治疗性蛋白质的哺乳动物细胞。这个问题将通过仔细的建模和实验来解决，包括温度的操纵，代谢物和代谢途径，生物反应器的操作模式，以及在所考虑的生物体的细胞周期中穿越的速度。一个重要的目标是培养精通生物过程优化的生物学和数学方面的研究生。最终，提出的工作目标是为设计基于哺乳动物细胞的治疗性蛋白质处理系统制定一个强大的策略，包括培养基设计，生物反应器操作条件和操作模式。要使用的一般策略可以定义为宏观代谢工程与细胞水平建模相结合。高度可再生的动物细胞培养系统的发展将对生物制药行业以外的许多生物工业应用产生重大影响。

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. The future of the biopharmaceutical industry will rely on the development and enhancement of these mammalian cell culture production systems. Such biologics are produced in relatively small quantities due to the highly specialized culture conditions needed and the susceptibility of the culture to either reduced productivity or significant cell death as a result of slight changes in culture conditions. While earlier generations of biochemical engineers focused on the non-biological aspects of production, such as reactor design, more recent engineering developments have recognized that the greatest potential for enhancing production lies within the animal cells themselves.In this proposal, an important and often neglected aspect of bioprocess development, namely bioprocess optimization is addressed. This proposal deals with the development of a novel optimization strategy for production of therapeutic proteins by mammalian cells. The problem will be addressed by careful modeling and experiments including the manipulation of temperature, metabolites and metabolic pathways, bioreactor operating mode, and rate of traverse through the cell cycle of the organism under consideration. An important goal is the training of graduate students who are well versed in both the biological and the mathematical aspects of bioprocess optimization.Ultimately, the objective of the proposed work is to develop a robust strategy for the design of mammalian cell based therapeutic protein processing systems, including media design, bioreactor operation conditions and operating mode. The general strategy to be used can be defined as macro-scale metabolic engineering combined with modeling at the cellular level. The development of highly reproducible animal cell culture systems will have significant impact for many bioindustrial applications beyond the biopharma industry.