Model-driven Media and Process Optimization in Mammalian Cell Lines

哺乳动物细胞系中模型驱动的培养基和过程优化

• 批准号: 7538942
• 负责人: IMAN FAMILI
• 金额: $ 87.29万
• 依托单位: GT LIFE SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7538942
• 关键词: Accounting; Adipocytes; Antibodies; Antibody Formation; Benchmarking; Biochemical Pathway; Biological Products; Biological Sciences; Bioreactors; Boxing; Case Study; Cell Culture System; Cell Density; Cell Line; Cells; Chemicals; Clinical; Collaborations; Computer Simulation; Condition; Cultured Cells; Custom; Data; Development; Drug Formulations; Economics; Engineering; Ensure; Equilibrium; Evaluation; Gene Proteins; Genome; Genomics; Goals; Growth; Healthcare; Human; Hybridomas; Immunoglobulin G; Mammalian Cell; Marketing; Measurement; Measures; Medicine; Metabolic; Metabolic Pathway; Metabolism; Methods; Modeling; Modification; Multiple Myeloma; Mus; Muscle Cells; Nutrient; Nutritional; Output; Parents; Patients; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Physiological; Physiology; Price; Procedures; Process; Production; Productivity; Proteins; Public Health; Publishing; Range; Rate; Reaction; Reagent; Recombinant Proteins; Reporting; Research; Research Activity; Research Project Grants; Sales; Screening procedure; Serum; Simulate; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Source; Standards of Weights and Measures; Supplementation; System; Techniques; Technology; Testing; Therapeutic; TimeLine; United States Food and Drug Administration; Validation; Vitamins; Work; base; bioprocess; cellular engineering; cofactor; commercial application; commercialization; computerized tools; concept; cost; design; feeding; improved; interest; models and simulation; next generation; novel therapeutics; programs; reconstruction; simulation; success; therapeutic protein; uptake; vector

DESCRIPTION (provided by applicant): Protein-based therapeutic products have contributed immensely to healthcare and constitute a large and growing percentage of the total pharmaceutical drugs. The majority of these FDA approved products are manufactured using mammalian cell culture systems. Over the past 10-20 years substantial progress has been made to overcome some of the key barriers to large-scale mammalian cell culture. Despite these improvements, the development of new biopharmaceutical products remains an expensive and lengthy process, where 20-30% of the total cost is associated with process development and clinical manufacturing. Currently most process optimization strategies are performed using a trial and error approach where cells are treated as a `black box' and process outputs are improved over several months by laborious 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, enabled by computational modeling and simulation technologies, can greatly improve and accelerate media and process development in mammalian cell line systems. In the Phase I of this SBIR research project, we utilized our computational modeling platform and expertise in metabolic modeling and mammalian cell culture to reduce byproduct formation in a GS-NS0 murine myeloma cell line in collaboration with SAFC (Sigma-Aldrich Fine Chemicals) Biosciences. To implement our model-driven media optimization approach, we reconstructed a metabolic model for NS0 cell containing 456 metabolites and 470 metabolic reactions. The model was used to develop nutritional modifications to the basal media to reduce byproduct formation and improve growth and productivity. Experimental evaluation of our model-based media formulations in NS0 cell culture showed significant improvements over traditional methods for media analysis and resulted in approximately 12% lower lactate and up to 67% higher final product titers. With the successful completion of our Phase I proof-of-concept study, we now put forth a Phase II proposal that aims at completing the development of a commercial platform for media and process optimization that significantly improves the existing timelines associated with therapeutic protein production in mammalian cell lines. We plan to achieve this goal through: (1) refinement and expansion of the metabolic model of NS0 cell line, (2) integration of a transient flux balance approach for quantitative implementation of media designs, and (3) validation of the final framework using three case studies for antibody production in NS0 cell line. We will measure the overall success in Phase II by our ability to reduce the timelines to develop an optimized media that results in lower byproduct formation and higher productivity in cell culture. Successful completion of the specific aims outlined in the proposed plan will benchmark the commercial value of a model-driven approach in recombinant protein production through rational selection of nutrient supplementation and process optimization strategies. PUBLIC HEALTH RELEVANCE: Protein-based therapeutic products have contributed immensely to healthcare and constitute a large and growing percentage of the total pharmaceutical market. The majority of these FDA approved products are manufactured using mammalian cell culture systems. The proposed work aims at developing computational strategies that significantly improves the timelines and cost associated with therapeutic protein production in mammalian cells. 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.

描述(申请人提供)：以蛋白质为基础的治疗产品对医疗保健做出了巨大贡献，在所有药物中占很大比例，而且比例还在不断增长。这些FDA批准的产品中的大多数都是使用哺乳动物细胞培养系统生产的。在过去的10-20年里，在克服大规模哺乳动物细胞培养的一些关键障碍方面取得了实质性进展。尽管有这些改进，新生物制药产品的开发仍然是一个昂贵和漫长的过程，总成本的20%-30%与工艺开发和临床制造有关。目前，大多数流程优化策略都是采用试错法进行的，将单元视为“黑匣子”，经过几个月的艰苦试验，流程产出得到改善。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理相互作用知之甚少。通过计算建模和模拟技术对细胞系生理学和新陈代谢有一个基本的了解，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的发展。在这个SBIR研究项目的第一阶段，我们利用我们在代谢建模和哺乳动物细胞培养方面的计算建模平台和专业知识，与SAFC(Sigma-Aldrich Fine Chemals)Biosciences合作，减少了GS-NS0小鼠骨髓瘤细胞系的副产品形成。为了实现我们的模型驱动的培养基优化方法，我们重建了包含456个代谢物和470个代谢反应的NS0细胞的代谢模型。该模型被用来对基础基质进行营养改良，以减少副产品的形成，并提高生长和生产力。在NS0细胞培养中对我们的基于模型的培养基配方进行的实验评估显示，与传统的培养基分析方法相比，我们的培养基配方有了显着的改进，乳酸降低了约12%，最终产品效价提高了67%。随着我们的第一阶段概念验证研究的成功完成，我们现在提出一项第二阶段的建议，旨在完成介质和工艺优化的商业平台的开发，显著改善与哺乳动物细胞系治疗性蛋白质生产相关的现有时间表。我们计划通过以下方式实现这一目标：(1)改进和扩展NS0细胞系的代谢模型，(2)整合用于定量实施介质设计的瞬时通量平衡方法，以及(3)使用NS0细胞系产生抗体的三个案例研究验证最终框架。我们将通过缩短开发优化培养基所需时间的能力来衡量第二阶段的总体成功，这种优化培养基能降低细胞培养中副产物的形成并提高生产率。成功完成拟议计划中概述的具体目标，将通过合理选择营养补充和工艺优化策略，对重组蛋白生产中的模型驱动方法的商业价值进行基准测试。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都是使用哺乳动物细胞培养系统生产的。这项拟议的工作旨在开发计算策略，显著改善与哺乳动物细胞治疗性蛋白质生产相关的时间线和成本。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。