An Integrated Computational and Experimental Platform for CHO-based Protein Produ

描述(由申请人提供)：基于蛋白质的治疗产品对医疗保健做出了巨大贡献，在所有药物中占很大比例，而且比例还在不断增长。目前，大多数生物制药产品是使用哺乳动物细胞系生产的，特别是中国仓鼠卵巢(CHO)细胞系，该细胞系生产的治疗蛋白占美国许可的所有治疗蛋白的70%以上。目前，大多数蛋白质生产的细胞和过程开发策略都是使用试错法执行的，这种方法通过实验努力地改进过程输出。这些经验优化技术被广泛使用，因为在大多数情况下，人们对影响宿主细胞系生长和蛋白质生产的潜在生物因素知之甚少。对细胞系生理和代谢的基本了解可以极大地加快和改进产品开发和制造的进程。为了响应开发CHO细胞模型的商业需求，并以我们在微生物和哺乳动物细胞代谢建模方面的经验为基础，我们启动了一个计划，以开发一个集成的计算和实验平台，用于合理设计和优化CHO细胞系的蛋白质生产。这个集成平台将我们先进的建模技术与成熟的实验技术相结合，以加快时间线并改进CHO表达系统中治疗性蛋白质的生产过程。为了证明在这项研究中使用计算/实验集成方法的优势，我们打算通过代谢网络重建来表征通常用于增强蛋白质表达的丁酸钠补充对CHO细胞新陈代谢的影响，并确定在不需要防止细胞凋亡(丁酸盐治疗的不良副作用)的情况下导致类似比生产率提高的替代策略。成功完成本提案中提出的具体目标将清楚地证明，开发一种在CHO细胞系中生产重组蛋白的合理方法在科学和技术上是可行的，该方法可用于：(A)对细胞系对环境和遗传变化的反应有基本的了解，以及(B)开发新的代谢干预措施以提高蛋白质产量。公共卫生相关性：以蛋白质为基础的治疗产品对医疗保健做出了巨大贡献，在整个药品市场中占据了很大的比例，而且比例还在不断增长。这些FDA批准的产品中的大多数目前是使用哺乳动物细胞培养系统生产的。然而，大多数细胞和工艺开发策略都是采用试错法进行的，这使得生物制药产品的开发成为一个昂贵而漫长的过程。为了优化整个过程开发的需要，需要使用一种合理的方法，将建模技术与现有的实验技术相结合，从根本上改变宿主细胞系的开发和治疗性蛋白质的生产方式。降低治疗性蛋白质开发和制造的成本将确保下一代药物的数量足以满足患者的需求，价格足够低，患者能够负担得起。

DESCRIPTION (provided by applicant): Abstract Protein-based therapeutic products have contributed immensely to healthcare and constitute a large and growing percentage of the total pharmaceutical drugs. The majority of the biopharmaceutical products are currently manufactured using mammalian cell lines, in particular the Chinese hamster ovary (CHO) cell line which accounts for the production of more than 70% of all US licensed therapeutic proteins. Currently, most cell and process development strategies for protein production are performed using a trial and error approach where process outputs are improved laboriously by experimentation. These empirical optimization techniques are widely used because in most cases little is known about the underlying biological factors that impact growth and protein production in the host cell lines. A fundamental understanding of cell line physiology and metabolism can greatly accelerate and improve the process of product development and manufacturing. In response to commercial demands for developing a CHO cell model and building upon our experience with metabolic modeling in microbes and mammalian cells, we have initiated a program to develop an integrated computational and experimental platform for rational design and optimization of protein production in CHO cell lines. This integrated platform combines our advanced modeling technologies with established experimental techniques to accelerate the timelines and to improve the process of therapeutic protein production in the CHO expression system. To demonstrate the advantage of utilizing an integrated computational/experimental approach in this study, we intend to characterize the effect of sodium butyrate supplementation, commonly used to enhance protein expression, on CHO cell metabolism using a metabolic network reconstruction and identify alternative strategies that result in similar improvements in specific productivity without the need to prevent apoptosis (an undesirable side effect of butyrate treatment). Successful completion of the specific aims set forth in this proposal will provide a clear demonstration of the scientific and technical feasibility of developing a rational approach for recombinant protein production in CHO cell lines that can be used to: (a) generate fundamental understanding for cell line response to environmental and genetic changes, and (b) develop novel metabolic interventions for improved protein production. PUBLIC HEALTH RELEVANCE: Protein-based therapeutic products have contributed immensely to health care and constitute a large and growing percentage of t he total pharmaceutical market. The majority of these FDA approved products are currently manufactured using mammalian cell culture systems. However, most cell and process development strategies are performed using a trial and error approach, making biopharmaceutical product development an expensive and lengthy process. The need to optimize the overall process development calls for the use of a rational approach that combines modeling technologies with established experimental techniques to fundamentally change the way host cell lines are developed and therapeutic proteins are produced. Reducing the cost of therapeutic protein development and manufacturing would ensure that the next generation of medicines can be created in amounts large enough to meet patients' needs and at a price low enough that patients can afford them.