Development of Methodology to Determine in Vivo Metabolic Rates in Mammalian Cells as a Tool to Evaluate and Improve Bioreactor Performance

迄今为止，哺乳动物细胞生物反应器性能的优化已经通过应用与培养基组成和生物反应器操作相关的容易观察到的细胞动力学的经验相关性来实现。虽然取得了一定程度的成功，但这种经验主义方法可能取得的成功是有限的。通过以增加生物反应器性能的方式选择性地改变细胞代谢途径的活性，可以获得更大程度的成功。然而，如果不知道这些途径的调控特征，或者更重要的是，如果它们甚至不能精确测量，就很难以合理的方式改变途径的活性。一种测量体内代谢活动的方法的发展，正如本提案所提出的，解决了这一限制。这种方法将为哺乳动物细胞技术人员提供一个工具，从一个不同的、更有知识的方向来接近过程优化。

To date, the optimization of mammalian cell bioreactor performance has been implemented via the application of empirical correlations relating easily observable cellular kinetics to medium composition and bioreactor operation. While moderately successful, there is a limit to the success that may be achieved with such an empirical approach. A greater degree of success maybe obtained by selectively altering the activity of the metabolic pathways of the cells in a manner that increases bioreactor performance. However, it is difficult to alter pathway activities in a rational fashion if the regulatory characteristics of these pathways are not known, or, more importantly, if they can not even be accurately measured. The development of a method to measure in vivo metabolic activities, as presented in this proposal, addresses this limitation. This method will provide a tool for mammalian cell technologists to approach process optimization from a different and more knowledgeable direction.