A Unique Automated Bioreactor for Rearing Aseptic Mosquitoes from Larvae to Adults to Support Manufacture of Sanaria PfSPZ Products

项目摘要： 根据世界卫生组织的数据，2018年2.28亿疟疾病例增加了~16% 死亡人数为40.5万人，与2015年相比并未大幅减少。这个 世界需要一种疟疾疫苗。萨那里亚恶性疟原虫子孢子(PfSPZ)疫苗已保护 在美国、德国、美国、德国和印度进行的五项临床试验中，受试者100%控制人类疟疾感染(CHMI) 坦桑尼亚和马里；保护成年人至少18个月，防止严重的、不同种类的肺泡炎传播 在布基纳法索的临床试验中，并在6个非洲和2个欧洲的临床试验中进行了评估 在5个月到65岁的年龄段，美国有5个国家和5个地点。Sanaria已转移到符合GMP的第三阶段 生产，并正在向Sanaria®PfSPZ疫苗的许可和商业化迈进。至 生产足够的PfSPZ疫苗以满足广泛的世界市场的需求，特别是在 非洲，在负担得起的价格下，制造业的规模必须大大增加，商品成本(齿轮) 减少了。Sanaria的PfSPZ疫苗包含无菌、纯化、冷冻保存的活PfSPZ疫苗，这些疫苗 由无菌饲养的斯氏按蚊制造。目前的方法是感染，无菌， 蚊子的生产严重依赖人力和大量昂贵的消耗品，以及 需要相当大的空间。这个应用程序的总体目标是设计、构建、测试和改进一个半 自动化蚊子生物反应器将使无菌、感染PfSPZ的成虫A. Stephensi和将感染PfSPZ的蚊子转移到准备好的受感染蚊子转移容器(IMTC) 用于辐照和解剖，显著降低了PfSPZ产品的上游制造成本。在.期间 在第二阶段SBIR中，我们将最终确定一个半自动、闭合系统的无菌生产系统 蚊子从卵到受感染的成虫都准备接受照射。具体目标是：1)重新设计 幼虫生物反应器，用于从卵到幼虫再到成蚊的无菌无菌培养；2)重新设计成虫 蚊子容器第1部分：从幼虫生物反应器直接转移成虫；3)重新设计成蚊 容器第2部分：半自动将受感染蚊子转移到受感染蚊子转移容器； 4)重新设计受感染的蚊子转移容器，以实现免提转移和重复使用；以及5)整合所有 并在PfSPZ挑战赛的工程运行中演示它们在制造中的实用性。发展中 这个生物反应器将导致Sanaria生产PfSPZ产品的能力发生翻天覆地的变化， Sanaria在第三阶段GMP合规性显著降低的情况下走上了稳健、大规模生产的轨道 成本。

PROJECT ABSTRACT: According to the World Health Organization, the 228 million cases of malaria in 2018 were an increase of ~16 million cases over 2015 while the number of deaths was 405,000, not substantially less than in 2015. The world needs a malaria vaccine. Sanaria’s Plasmodium falciparum sporozoite (PfSPZ) vaccines have protected 100% of subjects against controlled human malaria infection (CHMI) in five clinical trials in the U.S., Germany, Tanzania, and Mali; protected adults for at least 18 months against intense, heterogeneous transmission of Pf in clinical trials in Burkina Faso, and have been assessed in clinical trials in 6 African and 2 European countries, and 5 sites in the US in 5 month to 65 year olds. Sanaria has moved to Phase 3 GMP compliant manufacturing, and is moving toward licensure and commercialization of Sanaria® PfSPZ Vaccine. To manufacture enough PfSPZ Vaccine to meet the needs of the extensive world markets, particularly those in Africa, at affordable prices, the scale of manufacturing must be greatly increased and cost of goods (COGs) reduced. Sanaria’s PfSPZ-based vaccines contain aseptic, purified, cryopreserved, live PfSPZ that are manufactured in aseptically reared Anopheles stephensi mosquitoes. The current method of infected, aseptic, mosquito production relies heavily upon human power and large quantities of expensive consumables, and requires significant space. The overarching aim of this application is to design, build, test and refine a semi- automated mosquito bioreactor, which will enable hands-free harvesting of aseptic, PfSPZ-infected, adult A. stephensi and transfer of PfSPZ-infected mosquitoes to an Infected Mosquito Transfer Container (IMTC) ready for irradiation and dissection, significantly reducing upstream manufacturing costs of PfSPZ products. During this Phase II SBIR we will finalize a system for semi-automated, closed-system production of aseptic mosquitoes from eggs through to infected adults ready for irradiation. The specific aims are: 1) Redesign the larval bioreactor for hands-free, aseptic culturing from eggs to larvae to adult mosquitoes; 2) Redesign adult mosquito container part 1: direct transfer of adults from the larval bioreactor; 3) Redesign adult mosquito container part 2: semi-automated transfer of infected mosquitoes to the Infected Mosquito Transfer Container; 4) Redesign the Infected Mosquito Transfer Container for hands free transfer and re-use; and 5) Integrate all steps and demonstrate their utility for manufacturing in an engineering run of PfSPZ Challenge. Developing this bioreactor will lead to a sea change in Sanaria’s capacity to manufacture its PfSPZ products, putting Sanaria on track for robust, mass-scale manufacturing at phase 3 GMP compliance at a significantly reduced cost.