Exploiting translation elongation for improved biologics manufacturing

利用平移伸长来改进生物制品的制造

• 批准号: 10760927
• 负责人: Shane Byrne
• 金额: $ 32.5万
• 依托单位: CODOMAX INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10760927
• 关键词: Accounting; Affect; Algorithms; Amino Acids; Antibodies; Area; Autoimmune Diseases; Biological; Biological Products; Biological Response Modifier Therapy; Biomanufacturing; Biotechnology; Case Study; Cell Line; Cells; Clinical; Codon Nucleotides; Complex; Development; Engineering; Family; Food; Foundations; Genes; Growth; Guanine + Cytosine Composition; Human; Industrialization; Industry; Interferon Alfa-2b; Interferon Type II; Interferon-beta; Interferons; Malignant Neoplasms; Marketing; Measures; Messenger RNA; Methods; Modeling; Modification; Organism; Pain; Pharmaceutical Preparations; Phase; Pichia; Pilot Projects; Process; Production; Protein Secretion; Proteins; RNA; Reagent; Recombinant Proteins; Resources; Running; Serum Albumin; Stress; Structure; System; Systems Biology; Technology; Therapeutic; Time; Transfer RNA; Translating; Translations; Validation; Virus Diseases; Work; biological adaptation to stress; cell type; commercialization; cost; cost effective; epitranscriptome; experience; glycosylation; improved; innovation; interferon therapy; mRNA Translation; manufacture; manufacturing capabilities; manufacturing run; off-patent; pointed protein; preservation; programs; protein complex; protein expression; research and development; response; therapeutic protein

Abstract – Codomax has developed a codon engineering platform that enables bespoke, high-yield, cost- effective protein manufacturing by leveraging a newly discovered translational regulatory system in industrially relevant cell systems. Protein manufacturing has a current estimated market size of over $300 billion, distributed among the areas of antibodies and biologic therapeutics, biotechnology, industrial protein reagents, and food. However, protein production is often a time-, cost- and resource-intensive process. Some proteins are difficult to express in high yields due to the inability of the cell to translate the gene sequences efficiently. Candidate proteins may be abandoned at the research and development phase due to an inability to produce them in sufficient yields for cost-effective commercial-scale production. Traditional codon optimization, a widely used strategy, can result in only modest yield improvements because it is grounded in incomplete and incorrect models of protein translation that assume a static pool of tRNAs within a given organism. Codomax has discovered that cells regulate translation in response to stress by altering the tRNA pool to match the codons enriched in stress response mRNAs, which in turn enhances translation of the codon-biased mRNAs. Codomax’s platform leverages this mechanism by using systems-level approaches to measure the codons that are most frequently translated during the production of a recombinant protein, accounting for a specific cell type under specific growth conditions and using this information to choose the truly optimal codons. This solution increases recombinant protein production in cells and enhances the translation of mRNA therapeutics, helping to meet current protein manufacturing demands. Pilot studies of this approach in Pichia pastoris, one of the three major cell lines used in protein manufacturing, resulted in a 2.4-fold increase in protein production compared to genes already optimized by existing commercial algorithms. In this Phase I project, Codomax will build on these findings by undertaking two specific aims: (1) optimizing codons for interferon production in P. pastoris and demonstrating an increase in protein yield of at least 200% as well as the ability to retain 90% functionality in IFN-α-2b and IFNγ, and (2) establishing the feasibility of the platform to enable complex protein production using N-glycosylated IFN-beta-1a and human serum albumin as case studies. Successful completion of these aims will both demonstrate proof-of-concept of our platform to produce commercially relevant therapeutic proteins in higher yields than existing technologies and demonstrate the feasibility of using P. pastoris for manufacturing complex biologics. Resonating well with the Biden Administration’s National Biotechnology and Biomanufacturing Initiative, this project provides a strong foundation for Phase II plans to scale our technology for commercialization, given the growing clinical utility of interferon therapies and the applicability of our technology to any type of biologic therapeutic. Our technology will ultimately find broad application in enabling development of biologics and lowering commercialization costs.

摘要 – Codomax 开发了一个密码子工程平台，可实现定制、高产、成本低的密码子工程平台。 利用新发现的翻译调控系统有效制造蛋白质 工业相关的细胞系统。蛋白质制造目前估计市场规模超过 300 美元 亿，分布于抗体及生物治疗、生物技术、工业蛋白等领域 试剂、食品。然而，蛋白质生产通常是一个时间、成本和资源密集型的过程。一些 由于细胞无法翻译基因序列，蛋白质难以高产表达 高效。候选蛋白可能由于无法在研究和开发阶段被放弃 以足够的产量生产它们，以实现具有成本效益的商业规模生产。传统密码子优化， 广泛使用的策略只能带来适度的产量提高，因为它基于不完整和 错误的蛋白质翻译模型假设给定生物体内有静态的 tRNA 库。科多马克斯有 发现细胞通过改变 tRNA 库以匹配密码子来调节翻译以应对压力 富含应激反应 mRNA，从而增强密码子偏倚 mRNA 的翻译。 Codomax 的平台利用这种机制，通过使用系统级方法来测量 在重组蛋白生产过程中最常翻译的密码子，会计 针对特定生长条件下的特定细胞类型，并使用此信息来选择真正的 最佳密码子。该解决方案增加了细胞中重组蛋白的产量并增强了翻译 mRNA 疗法，有助于满足当前的蛋白质制造需求。这种方法的试点研究 毕赤酵母是用于蛋白质制造的三种主要细胞系之一，其产量增加了 2.4 倍。 蛋白质产量与已通过现有商业算法优化的基因进行比较。在这个第一阶段 项目中，Codomax 将通过实现两个具体目标来建立这些发现：（1）优化密码子 巴斯德毕赤酵母中干扰素的产生，并且蛋白质产量增加了至少 200% 保留 IFN-α-2b 和 IFNγ 90% 功能的能力，以及 (2) 建立该平台的可行性 使用 N-糖基化 IFN-β-1a 和人血清白蛋白作为案例研究，能够生产复杂的蛋白质。 成功完成这些目标都将证明我们的平台的概念验证 商业相关的治疗性蛋白质的产量比现有技术更高，并证明了 使用巴斯德毕赤酵母制造复杂生物制剂的可行性。与拜登产生良好共鸣 政府的国家生物技术和生物制造计划，该项目提供了坚实的基础 鉴于干扰素的临床用途不断增长，第二阶段计划扩大我们的技术商业化规模 疗法以及我们的技术对任何类型的生物治疗的适用性。我们的技术最终将 在促进生物制品的开发和降低商业化成本方面有广泛的应用。