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.

项目摘要/摘要：疟疾的年度估计燃烧已增加到21600万个临床发作 仅在非洲，就有445,000人死亡，每年损失$ 12B的GDP。世界需要一个 疟疾疫苗。 Sanaria的恶性疟原虫孢子岩（PFSPZ）疫苗已保护100％ 在美国，德国的五项临床试验中，针对受控人类疟疾感染（CHMI）的受试者， 坦桑尼亚和马里；保护成年人至少6个月免受强烈的，异质的PF传播 两项在马里进行的临床试验，现在正在5个月内在临床试验中进行评估，到65岁的5名非洲人 国家和成年人在2个欧洲国家以及美国的3个地点。该计划是启动第一个 第三阶段临床试验，以支持向美国FDA提交生物制剂许可申请的提交 2018年下半年以及2019年上半年的非洲。Sanaria的基于PFSPZ的疫苗含有无菌性的疫苗， 纯化，冷冻保存，活疟原虫恶性疟原虫（PF）孢子岩（SPZ）（SPZ） 无菌饲养的围绕蚊子蚊子。当前无菌蚊子生产的方法，而 强大而可重复的，很大程度上依赖人力和大量昂贵的消耗品，以及 需要大量空间。蚊子解剖的半自动化以去除在下面开发的唾液网格 预计另一项SBIR赠款将使该过程的效率提高到每小时750-1,000个蚊子 运营商在2018年上半年。那个蚊子的生产现在是瓶颈，最昂贵的 踏上PFSPZ疫苗的GMP生产。因此，该项目的总体目的是发展 通过开发半降低人力和消耗成本和整体空间需求的方法 用于培养（制造）无菌蚊子的自动化系统。在特定目标1中，我们将设计，构建 并测试生物反应器的免提，无菌聚类，从卵到幼虫到成年蚊子。这 生物反应器将开发为1）满足所有GMP要求，2）消除对昂贵消耗品的需求 要增加产出，3）降低人力成本，4）消除了许多生物安全柜的需求 用于蚊子生产中，5）适用于从蚊子中去除孵化器的事件 生产。我们将证明新的生物反应器能够维持关键的无菌性 幼虫培养物的环境，并且在幼虫生物反应器中饲养的蚊子表现出等效性 与当前方法产生的那些。在特定目标2中，我们将开发自动化的设备 收集成年蚊子。我们的目的是能够直接转移新出现的成年蚊子 并从幼虫生物反应器中放入大型无菌蚊子容器中，蚊子将是 在配子细胞上喂养并保持直到从蚊子中提取PFSPZ的那天。开发 幼体生物反应器将导致Sanaria生产其PFSPZ产品的能力，并放置 Sanaria在第3阶段GMP合规性的大规模制造业中，成本大大降低。