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.

圣甲虫基因组学建立了一个独特的，微小的占地面积，扩展的细菌发酵过程它减少了基因组大肠杆菌，用于生产蛋白质治疗，尤其是疫苗结合物。过程将大大降低成本并升级这些关键疫苗成分的质量。此阶段IIB 重新提出的重点是Scarab首款疫苗产品的商业开发，即白喉突变体 Toxin CRM197是非常成功的共轭疫苗的重要组成部分，具有重大关注 NIH的任务。 CRM197还表现出了作为抗癌剂的希望。全球需求量很高，这是很难通过批处理技术生产，因此供应受到限制和不可靠。圣甲虫进入需要疫苗市场，CGMP级产品。为了提高效率，圣甲虫提议将最初的外包将CGMP级CRM197生产给一家将在其GMP设施中使用Scarab的创新流程的公司验证GMP制造的过程。在上一阶段2 SBIR项目中，我们的过程生成了高水平的纯重组CRM197，并产生始终高质量的固体和稳定的CRM197 蛋白质。特别是，A和B分解产品是Scarab高质量产品的次要组成部分。第2阶段的下游过程（DSP）导致高度纯CRM197现在在研究中出售市场。自最初的PH2B提案以来，新的研究已建立（i）成功缩放到10升生产性发酵过程以及（ii）动态更现代，更有效的DSP，可自动化，一个商业生存能力的重要要求。 Waisman生物制造（WB），一个良好的非营利性CGMP合同开发和制造组织将执行CGMP级CRM197的生产。选择WB是因为它具有提供了非常有竞争力的报价，并且具有出色的经验和专业知识 - 它已经制造了（并且它的客户发布了300多种临床产品，并提供了5-10个调查新药应用每年由他们的合作伙伴提交。 WB的位置与圣甲虫在同一城市中将有助于技术转移和项目监督。具体目的是： 1）新DSP的全面开发，扩展和自动化。发酵后期发展圣甲虫的过程，包括四种试点发酵和下游处理，技术转移到WB，和CGMP流程开发。 2）工程GMP原型运行，纯度和建立了效力。 3）三个GMP生产运行，可以在运行内和纯化后配方规格。 4）最终产品验证将包括纯度和效力分析（共轭和人类免疫原性）。 Nitto Avecia的稳定性研究。 GMP工程和生产运行的材料将为潜在客户提供评估样本。项目信息将用于生物大师在FDA上归档。