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

独特的自动化生物反应器，用于培育从幼虫到成虫的无菌蚊子，以支持 Sanaria PfSPZ 产品的生产。

• 批准号: 9620426
• 负责人: Peter F. Billingsley
• 金额: $ 30万
• 依托单位: SANARIA, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-18 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9620426
• 关键词: Adult; Africa; African; Air; Anopheles Genus; Attenuated; Automation; Biological; Biological Assay; Bioreactors; Cessation of life; Characteristics; Chemoprophylaxis; Child; Clinical; Clinical Trials; Country; Culicidae; Custom; Cyclic GMP; Disease; Dissection; Ebola virus; Ensure; Environment; Epidemic; European; Excision; Falciparum Malaria; Geographic Locations; Germany; Grant; Growth; Hand; Harvest; Heating; Hour; Human; Human Resources; Immunity; Incubators; Individual; Infection; Infection prevention; Institutes; Larva; Licensing; Licensure; Malaria; Malaria Vaccines; Mali; Manuals; Mass Vaccinations; Medical; Methods; Nutrient; Output; Parasites; Phase; Phase III Clinical Trials; Plasmodium falciparum; Plasmodium falciparum vaccine; Prevention; Price; Process; Production; Pupa; Radiation; Reporting; Reproducibility; Risk; Safety; Salivary Glands; Sea; Shapes; Site; Small Business Innovation Research Grant; Sporozoite vaccine; Sporozoites; Stream; System; Tanzania; Testing; Tube; Vaccines; Waste Products; base; commercialization; cost; design; egg; fitness; flasks; human old age (65+); improved; malaria infection; manufacturing process; protective efficacy; scale up; seal; success; tool; transmission process

Project Summary/Abstract: The annual estimated burden of malaria has increased to 216M clinical episodes and 445,000 deaths, and annual losses of >$12B of GDP to malaria in Africa alone. 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 at least for 6 months against intense, heterogeneous transmission of Pf in two clinical trials in Mali, and are now being assessed in clinical trials in 5 months to 65-year old’s in 5 African countries and in adults in 2 European countries as well as 3 sites in the US. The plan is to initiate the first Phase 3 clinical trials to support submission of a Biologics License Application to the FDA in the US in the second half of 2018, and in Africa in the first half of 2019. Sanaria’s PfSPZ-based vaccines contain aseptic, purified, cryopreserved, live Plasmodium falciparum (Pf) sporozoites (SPZ) that are manufactured in aseptically reared Anopheles stephensi mosquitoes. The current method of aseptic mosquito production, while robust and reproducible, relies heavily upon manpower and large quantities of expensive consumables, and requires significant space. Semi-automation of mosquito dissection to remove salivary glands developed under another SBIR grant is expected to increase the efficiency of the process to 750-1,000 mosquitoes per hour per operator in the first half of 2018. Thus, production of mosquitoes is now the bottleneck and most expensive step in the GMP manufacture of PfSPZ vaccines. The overarching aim of this project is therefore to develop methods to reduce manpower and consumable costs and overall space requirements by developing a semi- automated system for culturing (manufacturing) aseptic mosquitoes. In specific aim 1, we will design, build and test a bioreactor for hands-free, aseptic culturing from eggs to larvae to adult mosquitoes. This bioreactor will be developed to 1) meet all GMP requirements, 2) remove the need for expensive consumables to increase output, 3) reduce human costs, 4) eliminate the need for many of the biological safety cabinets used in mosquito production, and 5) be adaptable for eventual removal of incubators from mosquito production. We will demonstrate that the new bioreactor is capable of maintaining the critical aseptic environment for the larval cultures, and that the mosquitoes raised in the larval bioreactor show equivalence with those produced by current methods. In specific aim 2 we will develop apparatus for automated harvesting of adult mosquitoes. Our aim is to be able to transfer newly emerged adult mosquitoes directly and hands free from the larval bioreactors into big aseptic mosquito containers in which the mosquitoes will be fed upon gametocytes and maintained until the day of extraction of PfSPZ from mosquitoes. Developing the larval bioreactor will lead to a sea change in Sanaria’s capacity to manufacture its PfSPZ products, putting Sanaria on track for mass-scale manufacturing at phase 3 GMP compliance at a significantly reduced cost.

项目概要/摘要：疟疾的年度估计负担已增加到2.16亿次临床发作 44.5万人死亡，仅在非洲，每年因疟疾造成的国内生产总值损失就超过120亿美元。世界需要一个 疟疾疫苗Sanaria的恶性疟原虫子孢子（PfSPZ）疫苗保护了100%的 受试者在美国的五项临床试验中对抗受控人类疟疾感染（CHMI），德国， 坦桑尼亚和马里;保护成年人至少6个月免受密集的，异质性的Pf传播， 在马里进行了两项临床试验，目前正在5个月内对5名非洲65岁以上的人进行临床试验评估。 在2个欧洲国家以及美国的3个研究中心的成人中。计划是启动第一个 3期临床试验，以支持在美国向FDA提交生物制品许可证申请， 2018年下半年，2019年上半年在非洲。Sanaria的PfSPZ疫苗含有无菌， 纯化、冷冻保存的活恶性疟原虫（Pf）子孢子（SPZ）， 无菌饲养的斯氏按蚊。目前的无菌蚊子生产方法， 坚固耐用且可重复，严重依赖人力和大量昂贵的耗材， 需要很大的空间。半自动化的蚊子解剖，以去除唾液腺， 另一笔SBIR赠款预计将把这一过程的效率提高到每小时750- 1000只蚊子， 2018年上半年的运营商。因此，蚊子的生产现在是瓶颈和最昂贵的 PfSPZ疫苗的GMP生产步骤。因此，该项目的总体目标是开发 通过开发一种半导体器件，降低人力和消耗品成本以及总体空间需求的方法， 用于培养（制造）无菌蚊子的自动化系统。在具体目标1中，我们将设计、建造 并测试一个生物反应器，用于从卵到幼虫再到成年蚊子的免提无菌培养。这 生物反应器将被开发为1）满足所有GMP要求，2）消除对昂贵消耗品的需求 增加产量，3）减少人力成本，4）消除对许多生物安全柜的需要， 用于蚊子的生产，以及5）适于最终从蚊子身上去除孵化器 生产我们将证明新的生物反应器能够保持关键的无菌条件， 幼虫培养的环境，以及在幼虫生物反应器中饲养的蚊子表现出等效性 与通过当前方法产生的那些相比。在具体目标2中，我们将开发用于自动化的设备。 捕获成年蚊子。我们的目标是能够直接转移新出现的成年蚊子 从幼虫生物反应器中解放出来，放入大的无菌蚊子容器中， 以配子母细胞喂养并维持直至从蚊子中提取PfSPZ的那天。发展 幼虫生物反应器将导致Sanaria生产PfSPZ产品的能力发生巨大变化， Sanaria正在以显著降低的成本进行第3阶段GMP合规的大规模生产。