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

在稳定的基因组减少的大肠杆菌菌株中产生类毒素佐剂 CRM197

基本信息

批准号：
9897524
负责人：
FREDERICK R BLATTNER
金额：
$ 100万
依托单位：
SCARAB GENOMICS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-05 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9897524
关键词：
Adjuvant Amino Acids Antibodies Antineoplastic Agents Automation Bacterial Meningitis Bacterial Pneumonia Biological Biological Assay Biological Products Biomanufacturing Carrier Proteins Cities Client Clinical Conjugate Vaccines Contracts Cyclic GMP Development Diphtheria Diphtheria Toxin Disease Documentation Endotoxins Engineering Escherichia coli Evaluation Fermentation Formulation Freezing Future Genome Genomics High Pressure Liquid Chromatography Human Industry Investigational New Drug Application Liquid substance Location Manufacturer Name Market Research Methods Minor Mission Modernization Phase Pneumonia Procedures Process Production Proteins Protocols documentation Public Health Quality Control Recombinants Reference Values Research Resources Running Sales Sampling Small Business Innovation Research Grant Speed System T-Lymphocyte Technology Technology Transfer Testing Time Toxin Toxoids Training United States National Institutes of Health Universities Vaccines Validation Wisconsin cGMP production cell bank cost cross reacting material 197 experience immunogenicity improved innovation interest manufacturing process microbial mutant operation prevent prototype scale up stability testing therapeutic protein vaccine development

项目摘要

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.

Scarab Genomics 建立了一种独特的、占地面积小、扩展的细菌发酵工艺，其基因组减少的大肠杆菌，用于生产蛋白质疗法，特别是疫苗缀合物。将大幅降低成本并提升这些关键疫苗成分的质量。重新提交的重点是圣甲虫第一个疫苗产品白喉突变体的商业开发毒素 CRM197 是非常成功的结合疫苗的重要组成部分，引起了人们的极大兴趣 CRM197 还显示出作为一种在全球范围内需求量很大的抗癌药物的前景。由于难以通过批量技术生产，因此供应受到限制且不可靠。疫苗市场需要 cGMP 级产品为了获得最大效率，Scarab 建议外包初始产品。为一家公司生产 cGMP 级 CRM197，该公司将在其 GMP 设施中使用 Scarab 的创新工艺，验证 GMP 制造流程在之前的第 2 阶段 SBIR 项目中，我们生成了流程。高水平的纯重组 CRM197 并持续产生高质量的可溶且稳定的 CRM197 特别是，A 和 B 分解产物是 Scarab 优质产品的次要成分。第 2 阶段的下游工艺 (DSP) 产生了高纯度的 CRM197，现已在研究中出售自最初的 Ph2B 提案以来，新的研究已经确定 (i) 成功扩大至 10L。高效的发酵过程和 (ii) 更加现代和高效的自动化 DSP 商业可行性的重要要求。 Waisman Biomanufacturing (WB) 是一家成熟的非营利性 cGMP 合同开发和制造组织将进行 cGMP 级 CRM197 的生产，因为它具有提供了非常有竞争力的报价，并拥有卓越的经验和专业知识 - 它已经制造（和其客户已发布）超过 300 种临床级产品，其中包括 5-10 个研究性新药申请每年由其合作伙伴提交的 WB 与 Scarab 位于同一城市将促进技术发展。转让和项目监督的具体目标是： 1）新型DSP发酵后期开发的全面开发、规模化和自动化。 Scarab 的工艺，包括四次试点发酵和下游加工、向 WB 的技术转让、和 cGMP 工艺开发 2) 工程 GMP 原型运行、有关纯度和规格的规范。 3) 三个 GMP 生产运行，制定运行内和运行后纯化 4) 最终产品验证，包括纯度和效力测定（结合和人类 Nitto Avecia 进行的稳定性研究来自 GMP 工程和生产运行。为潜在客户提供样品进行评估项目信息将用于生物大师。向FDA备案。