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

Scarab Genomics建立了一种独特的、占地面积小的、延长的细菌发酵过程， 其简化的基因组E.大肠杆菌，用于生产蛋白质治疗剂，特别是疫苗缀合物。过程 将大大降低成本并提升这些关键疫苗成分的质量。本阶段IIB 重新提交的重点是圣甲虫的第一个疫苗产品，白喉突变体的商业开发 毒素CRM 197是非常成功的缀合疫苗的重要组分， NIH的使命。CRM 197也显示出作为抗癌剂的前景。在全球范围内需求量很大， 难以通过批量技术生产，因此供应受到限制且不可靠。为了让圣甲虫进入 疫苗市场，需要cGMP级产品。为了最大限度地提高效率，Scarab建议将最初的 生产cGMP级CRM 197的公司将在其GMP设施中使用圣甲虫的创新工艺， 验证GMP生产工艺。在之前的第2阶段SBIR项目中，我们的流程生成了 更高水平的纯重组CRM 197，并产生一致的高质量可溶性和稳定的CRM 197 蛋白特别是，A和B分解产物是圣甲虫高质量产品的次要成分。 第2阶段的下游工艺（DSP）产生了高纯度的CRM 197，现在在研究中销售 市场自最初的Ph 2B提案以来，新的研究已经建立了（i）成功的规模高达10 L 生产发酵过程和（ii）一个显着更现代化和有效的DSP是自动化的， 这是商业可行性的重要要求。 魏斯曼生物制造（WB），一个完善的非营利cGMP合同开发和 生产组织将生产cGMP级CRM 197。WB被选中是因为它有 提供了一个非常有竞争力的报价，并具有特殊的经验和专业知识-它已经制造（和 其客户已发布）超过300种临床级产品，5-10种研究性新药申请 每年都由他们的合作伙伴提交。WB与Scarab位于同一城市， 转让和项目监督。具体目标是： 1)新DSP的全面开发、规模扩大和自动化。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，有关纯度和 建立的力量。3)三次GMP生产运行，在运行内和纯化后配制 规范. 4)最终产品验证，包括纯度和效价测定（结合和人 免疫原性）。Nitto Avidon的稳定性研究。来自GMP工程和生产运行的材料将 为潜在客户提供样品进行评估。项目信息将用于生物硕士 在FDA备案。

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.