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

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.