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，成本- 通过利用新发现的翻译调节系统， 工业上相关的细胞系统。蛋白质制造业目前的市场规模估计超过300美元。 亿美元，分布在抗体和生物治疗，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 由于细胞不能翻译基因序列， 有效地候选蛋白质可能在研究和开发阶段被放弃，因为它们不能 以足够的产量生产它们，以进行具有成本效益的商业规模生产。传统的密码子优化， 这是一种广泛使用的策略，只能带来适度的产量提高，因为它是基于不完整和 错误的蛋白质翻译模型，假设给定生物体内的tRNA是静态的。Codomax拥有 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，这反过来又增强了密码子偏向mRNA的翻译。 Codomax的平台通过使用系统级方法来衡量 在重组蛋白生产过程中最常翻译的密码子，占 在特定生长条件下的特定细胞类型，并使用这些信息来选择真正的 最佳密码子这种解决方案增加了细胞中重组蛋白的产量，并增强了蛋白质的翻译。 mRNA疗法，有助于满足当前的蛋白质制造需求。对这一办法的试点研究， 巴斯德毕赤酵母是蛋白质生产中使用的三种主要细胞系之一， 与已经通过现有商业算法优化的基因相比，蛋白质产量。这项I期 Codomax项目将在这些发现的基础上实现两个具体目标：（1）优化密码子， 在巴斯德毕赤酵母中产生干扰素，并证明蛋白质产量增加至少200%，以及 在IFN-α-2b和IFNγ中保留90%功能性的能力，以及（2）建立平台的可行性， 能够使用N-糖基化IFN-β-1a和人血清白蛋白作为案例研究生产复杂蛋白质。 成功完成这些目标将证明我们的平台概念验证， 商业上相关的治疗性蛋白质的产量高于现有技术，并证明了 利用巴斯德毕赤酵母生产复杂生物制剂的可行性。与拜登产生共鸣 美国政府的国家生物技术和生物制造倡议，该项目提供了一个坚实的基础 考虑到干扰素临床应用的不断增长， 治疗和我们的技术适用于任何类型的生物治疗。我们的技术将最终 在促进生物制品开发和降低商业化成本方面具有广泛的应用。

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.