Exploiting translation elongation for improved biologics manufacturing

摘要-Codomax开发了一个密码子工程平台，使定制、高产量、低成本- 通过利用新发现的翻译调控系统有效地制造蛋白质 与工业相关的电池系统。蛋白质制造目前的市场规模估计超过300美元。 10亿美元，分布在抗体和生物疗法、生物技术、工业蛋白质等领域 试剂和食物。然而，蛋白质生产往往是一个时间、成本和资源密集型的过程。一些人 由于细胞无法翻译基因序列，蛋白质很难高产量地表达 效率很高。候选蛋白质在研究和开发阶段可能会因为不能 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。传统的密码子优化， 一种广泛使用的策略，只能带来适度的产量提高，因为它植根于不完整和 错误的蛋白质翻译模型，假设给定生物体内有一个静态的tRNA池。Codomax有 发现细胞通过改变tRNA池来匹配密码子来调节翻译以应对压力 丰富的应激反应mRNAs，这反过来又增强了偏向密码子的mRNAs的翻译。 CoDomax的平台利用这一机制，使用系统级的方法来衡量 在重组蛋白生产过程中最频繁翻译的密码子，占 在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。这种溶液增加了细胞中重组蛋白的产量，并增强了翻译 信使核糖核酸疗法，帮助满足目前的蛋白质制造需求。对此方法的试点研究 毕赤氏酵母是用于蛋白质制造的三个主要细胞系之一，其结果是 将蛋白质生产与已被现有商业算法优化的基因进行比较。在此阶段I 项目，Codomax将以这些发现为基础，承担两个具体目标：(1)优化密码子以 在巴斯德毕赤酵母中生产干扰素，蛋白质产量至少增加200%， 在干扰素-α-2b和干扰素γ中保留90%功能的能力，以及(2)建立该平台的可行性 以N-糖基化干扰素-β-1a和人血清白蛋白为例，实现复合蛋白的生产。 成功完成这些目标将证明我们的平台将产生 与现有技术相比，具有商业意义的治疗性蛋白质的产量更高，并证明 巴斯德毕赤酵母用于生产复杂生物制品的可行性。与拜登产生很好的共鸣 政府的国家生物技术和生物制造倡议，该项目提供了坚实的基础 考虑到干扰素的临床效用不断增长，For II计划将我们的技术扩展到商业化 治疗方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 在促进生物制剂的开发和降低商业化成本方面得到了广泛的应用。

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.