Model-driven Media Optimization in Hybridoma Cell Line

项目概述/摘要：基于蛋白质的治疗药物对医疗保健做出了巨大贡献，在药物总量中所占的比例越来越大。目前大多数生物制药产品是使用哺乳动物细胞系制造的。然而，与微生物系统相比，哺乳动物细胞培养物的生产力和产品滴度较低，主要是由于标准哺乳动物细胞培养物的生物量浓度较低。此外，哺乳动物细胞系会产生大量的废物，如乳酸、丙氨酸和氨，这些废物会降低生物量和蛋白质的产量，导致毒性调节，并抑制细胞生长。目前，哺乳动物细胞培养的培养基和工艺优化策略是使用试错方法进行的，其中通过实验费力地改进了工艺输出。这些经验优化技术被广泛使用，因为在大多数情况下，对影响宿主细胞系生长和蛋白质产生的潜在生理相互作用知之甚少。对细胞系生理和代谢的基本理解可以极大地改善和加速哺乳动物细胞系系统的培养基和过程发育。在培养基优化和工艺开发中，合理的设计方法需要能够捕获和分析宿主细胞系潜在生理学的建模和仿真技术。在这个SBIR研究计划中，我们打算通过重建小鼠杂交瘤细胞系模型来证明代谢建模在培养基优化中的价值。结合我们的建模技术和使用重建的杂交瘤代谢模型，我们将设计硅片衍生的培养基配方，以减少杂交瘤细胞培养中的副产物形成，并随后使用实验测量评估我们的培养基设计。该提案的成功将证明，通过合理选择营养补充策略，模型驱动改进重组蛋白生产的科学和技术可行性，并最终降低治疗性蛋白质开发和制造的成本，以确保下一代药物能够以足够大的数量满足患者的需求，并以足够低的价格使患者能够负担得起。项目说明：以蛋白质为基础的治疗药物对卫生保健作出了巨大贡献，在药物总量中所占的比例很大，而且还在不断增长。该提案的成功将证明，通过合理选择营养补充策略，模型驱动改进重组蛋白生产的科学和技术可行性，并最终降低治疗性蛋白质开发和制造的成本，以确保下一代药物能够以足够大的数量满足患者的需求，并以足够低的价格使患者能够负担得起。

DESCRIPTION (provided by applicant): Project Summary/Abstract: Protein-based therapeutics have contributed immensely to health care 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. However, compared to microbial systems, productivity and product titers in mammalian cell cultures are low mainly due to the low biomass concentration achievable in standard mammalian cell culture. In addition, mammalian cell lines produce significant amounts of waste products such as lactate, alanine, and ammonia, which reduce biomass yield and protein production, cause toxic accommodation, and inhibit cell growth. Currently, media and process optimization strategies in mammalian cell culture 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 physiological interactions that impact growth and protein production in the host cell lines. A fundamental understanding of cell line physiology and metabolism can greatly improve and accelerate media and process development in mammalian cell line systems. A rational design approach in media optimization and process development requires modeling and simulation technologies capable of capturing and analyzing the underlying physiology of the host cell line. In this SBIR research plan, we intend to demonstrate the value of metabolic modeling in media optimization using a reconstructed model of murine hybridoma cell line. In combination with our modeling technology and using the reconstructed model of hybridoma metabolism, we will design in silico-derived media formulations that reduce byproduct formation in hybridoma cell culture and subsequently evaluate our media designs using experimental measurements. Success of this proposal will demonstrate the scientific and technical feasibility of model-driven improvements in recombinant protein production through the rational selection of nutrient supplementation strategies and ultimately reduce the cost of therapeutic protein development and manufacturing to 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. Project Narrative: Protein-based therapeutics have contributed immensely to health care and constitute a large and growing percentage of the total pharmaceutical drugs. Success of this proposal will demonstrate the scientific and technical feasibility of model-driven improvements in recombinant protein production through the rational selection of nutrient supplementation strategies and ultimately reduce the cost of therapeutic protein development and manufacturing to 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.