Toxoid adjuvant CRM197 production in a stable reduced genome E. coli strain

圣甲虫基因组公司已经建立了一种独特的、占地面积很小的、延长的细菌发酵过程，通过 其还原基因组的大肠杆菌，用于生产蛋白质治疗药物，特别是疫苗结合物。这个过程 将大幅降低成本并提高这些关键疫苗组件的质量。此阶段IIB 重新提交的重点是Scarab的第一个疫苗产品-白喉突变株的商业开发 CRM197毒素，是非常成功的结合疫苗的重要组成部分，引起了 美国国立卫生研究院的使命。CRM197也显示出作为抗癌药物的前景。在全球范围内需求旺盛，它是 很难通过批量技术生产，因此供应受限且不可靠。为了让Scarab进入 疫苗市场上，cGMP级产品是必需的。为了最大限度地提高效率，Scarab建议将初始 将cGMP级CRM197的生产交给一家公司，该公司将在其GMP设施中使用Scarab的创新工艺， 对GMP生产工艺进行验证。在之前的第2阶段SBIR项目中，我们的流程生成了 高水平的纯重组CRM197，并产生稳定的优质可溶性CRM197 蛋白。特别是，A和B细分产品是Scarab高质量产品的次要组件。 第二阶段的下游工艺(DSP)产生了高纯度的CRM197，现在该研究已售出 市场。自从最初的Ph2B提案以来，新的研究已经建立了：(I)成功地扩展到10L 生产发酵过程和(Ii)显著更现代化和更高效的可自动化的数字信号处理器， 对商业可行性的至关重要的要求。 韦斯曼生物制造(WB)，一家久负盛名的非营利性cGMP合同开发和 制造组织，将执行cGMP级CRM197的生产。WB被选中是因为它拥有 提供极具竞争力的报价，并拥有非凡的经验和专业知识-它已制造(和 它的客户已经发布了300多个临床级产品，有5-10个正在研究的新药申请 由他们的合作伙伴每年提交。WB与Scarab位于同一城市将促进技术 转让和项目监督。具体目标是： 1)新一代数字信号处理器的全面开发、规模化和自动化。发酵的后期发展 Scarab的工艺，包括四个中试发酵和下游加工，向WB转让技术， 和cGMP工艺开发。2)工程GMP原型运行，关于纯度和 效力已建立。3)三个GMP生产流程，规定生产过程中和净化后 规格。4)最终产品验证，将包括纯度和效力分析(接合和人 免疫原性)。稳定性研究：Nitto Avecia。来自GMP工程和生产运行的材料将 为潜在客户提供样品以供评估。项目信息将在生物大师中使用 向FDA提交文件。

Scarab Genomics has established a unique, tiny footprint, extended bacterial fermentation process enabled by its reduced genome E. coli, for production of protein therapeutics, especially vaccine conjugates. The process will massively reduce the cost and upgrade the quality of these critical vaccine components. This Phase IIB resubmission focuses on commercial development of Scarab’s first vaccine product, the diphtheria mutant toxin CRM197, an important component of highly successful conjugate vaccines, of significant interest to the mission of the NIH. CRM197 has also shown promise as an anti-cancer agent. In high demand worldwide, it is difficult to produce by batch technology, so supply is constrained and unreliable. For Scarab to enter the vaccine market, cGMP-grade product is required. For maximum efficiency, Scarab proposes to outsource initial production of cGMP-grade CRM197 to a company that will use Scarab’s innovative process in its GMP facility, validating the process for GMP manufacturing. In a previous Phase 2 SBIR project, our process generated high levels of pure recombinant CRM197 and yielded consistently high quality soluble and stable CRM197 protein. In particular, the A and B breakdown products are minor components of Scarab’s high-quality product. The downstream process (DSP) from Phase 2 resulted in highly pure CRM197 now sold on the research market. Since the original Ph2B proposal, new research has established (i) successful scale up to a 10L productive fermentation process and (ii) a dramatically more modern and efficient DSP that is automatable, a vital requirement for commercial viability. Waisman Biomanufacturing (WB), a well-established not-for-profit cGMP contract development and manufacturing organization, will perform production of cGMP-grade CRM197. WB was selected because it has provided a very competitive quote, and has exceptional experience and expertise – it has manufactured (and its clients have released) well over 300 clinical-grade products, with 5-10 investigational new drug applications submitted by their partners each year. WB’s location in the same city as Scarab will facilitate technology transfer and project oversight. The Specific Aims are: 1) Full development, scale-up and automation of the new DSP. Late-stage development of the fermentation process at Scarab, including four pilot fermentations and downstream processing, technology transfer to WB, and cGMP process development. 2) engineering GMP prototype run, specifications regarding purity and potency established. 3) Three GMP production runs that formulize within-run and post-purification specifications. 4) Final product validation that will include purity and potency assays (conjugation and human immunogenicity). Stability studies by Nitto Avecia. Materials from GMP engineering and production runs will provide potential customers with samples for evaluation. Project information will be used in a Biological Master File at the FDA.