Modularizing manufacture of PfSPZ vaccines: ookinete production for PfSPZ manufacture in mosquitoes and in vitro

PfSPZ 疫苗的模块化生产：在蚊子和体外生产 PfSPZ 的动合生产

基本信息

批准号：
10761373
负责人：
STEPHEN Lev HOFFMAN
金额：
$ 28.22万
依托单位：
SANARIA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-06 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10761373
关键词：
Address African Anopheles Genus Antigens Biological Biological Assay Bioreactors Blood Chitinase Complex Country Cryopreservation Culicidae Dissociation Divorce Excision Female Fluorescent Dyes Future Germ Cells Goals Good Manufacturing Process Grant Human In Vitro Infection Link Location Magnetism Malaria Malaria Vaccines Membrane Membrane Proteins Methods Nature Oocysts Parasites Parasitic infection Plasmodium falciparum Plasmodium falciparum vaccine Pregnant Women Process Production Protocols documentation Quality Control Readiness Serum Specific qualifier value Sporozoite vaccine Sporozoites Standardization Structure Survival Rate Testing Time Vaccination Vaccine Production Vaccines Work cell motility comparative cost density extracellular feeding flexibility large scale production malaria infection malaria transmission male manufacture matrigel prevent timeline transmission process uptake vaccine efficacy zygote

Address African Anopheles Genus Antigens Biological Biological Assay Bioreactors Blood Chitinase Complex Country Cryopreservation Culicidae Dissociation Divorce Excision Female Fluorescent Dyes Future Germ Cells Goals Good Manufacturing Process Grant Human In Vitro Infection Link Location Magnetism Malaria Malaria Vaccines Membrane Membrane Proteins Methods Nature Oocysts Parasites Parasitic infection Plasmodium falciparum Plasmodium falciparum vaccine Pregnant Women Process Production Protocols documentation Quality Control Readiness Serum Specific qualifier value Sporozoite vaccine Sporozoites Standardization Structure Survival Rate Testing Time Vaccination Vaccine Production Vaccines Work cell motility comparative cost density extracellular feeding flexibility large scale production malaria infection malaria transmission male manufacture matrigel prevent timeline transmission process uptake vaccine efficacy zygote

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项目摘要

In 2022 WHO called for highly efficacious vaccines against malaria that prevent Plasmodium falciparum (Pf) infection in > 90% of recipients. Alone among malaria vaccines, Sanaria® PfSPZ (Pf sporozoite (SPZ)) vaccines have shown > 90% vaccine efficacy (VE) against controlled human malaria infection and VE against intense field transmission of malaria to pregnant women for two transmission years without boosting. Our goals are to further increase potency, increase efficiency and consistency of manufacture and reduce cost of goods (COGS) to be able to use a PfSPZ vaccine for elimination of malaria. PfSPZ vaccines are manufactured using stage V Pf gametocytes fed to mosquitoes, a process that includes tight coordination between culture of gametocytes and mosquito rearing. Most importantly, Sanaria has developed methods for producing PfSPZ without mosquitoes in bioreactors (Nature, 2022). Transitioning to bioreactor production would increase efficiency of manufacture by at least 10-fold, reduce COGS by >90%, and allow for PfSPZ vaccine production worldwide. This proposal is intended to provide mass-produced, aseptic, purified, cryopreserved ookinetes that can be used to increase efficiency of production of mosquito- and bioreactor-produced PfSPZ vaccines. We will optimize production, purification, cryopreservation, QC release, and use in manufacturing of ookinetes. The early steps in manufacturing that end with ookinetes can then be de- linked from the later steps that commence with ookinetes, allowing both to proceed separately, unconstrained by timing or physical location. The work will be addressed in 4 Specific Aims. 1: Optimize ookinete production. Media and culture conditions for stage V gametocytes will be optimized for conversion to ookinetes. Readouts will include female:male ratio, macrogametocyte density, and stage specific antigen expression. Our target is a method that consistently results in 25% conversion of stage V gametocytes to ookinetes. 2: Develop and optimize methods for ookinete purification. We will test multiple methods, selecting those that can be scaled. Our target is Pf ookinetes that are 100% free of RBCs with a yield of 80% using a method that can be adapted to large scale manufacturing. 3: Develop a protocol for ookinete cryopreservation. Sanaria routinely cryopreserves PfSPZ with consistent, excellent results. Ookinetes are similar to PfSPZ. Both are motile, have a multimembrane pellicular complex, express a dominant membrane protein, and are similar in size. A range of cryoprotectant additives (CPA), cooling and warming rates, and CPA dilution methods will be tested. Our target is a survival rate of at least 50%. 4: Develop assays for ookinete viability and potency. 4 approaches will be tested: a) Membrane integrity, b) Motility, c) Infectivity to mosquitoes to produce PfSPZ, and d) In vitro conversion of ookinetes to 3- and 8-day oocysts and iPfSPZ.

在2022年，他要求对疟疾进行高效疫苗，以防止疟原虫 > 90％的受体中的恶性疾病（PF）感染。 Sanaria®PFSPZ（PF）单独使用疟疾疫苗 Sporozoite（SPZ））疫苗已显示针对人类受控的疫苗效率（VE）> 90％疟疾感染和反对疟疾向孕妇的强化现场传播的两种传输年没有提升。我们的目标是进一步提高效力，提高效率和制造的一致性并降低商品成本（COGS），以便能够使用PFSPZ疫苗消除疟疾。 PFSPZ疫苗是使用F级PF配子细胞制造的蚊子，包括配子细胞和蚊子文化之间的紧密协调的过程饲养。最重要的是，Sanaria开发了生产无蚊子的PFSPZ的方法在生物反应器中（自然，2022年）。过渡到生物反应器的生产将提高效率生产至少10倍，将COG降低> 90％，并允许PFSPZ疫苗生产全世界。该建议旨在提供批量生产，无菌，纯化，冷冻保存可用于提高蚊子和生物反应器生产的生产效率的Ookines PFSPZ疫苗。我们将优化生产，纯化，冷冻保存，QC发布和使用螺栓的制造。然后，可以使用邮轮结束的制造早期步骤链接从载量开始的后来步骤链接，允许两者单独进行，不受计时或物理位置的约束。这项工作将以4个特定目标解决。 1：优化Okinete生产。 V阶段的媒体和文化条件将优化用于转换为邮轮。读数将包括女性：男性比例，大胶质细胞密度和阶段特异性抗原表达。我们的目标是一种始终导致25％转化的方法第五阶段的配子细胞到邮轮。 2：开发并优化用于纯化的方法。我们将测试多种方法，选择可以缩放的方法。我们的目标是PF 100％使用可以大规模调整的方法的RBC，其产量为80％制造业。 3：制定用于冷冻保存的协议。 Sanaria通常是冷冻保存 PFSPZ具有一致，出色的结果。 Ookinetes与PFSPZ相似。两者都有动力，有一个多膜细胞复合物，表达主要的膜蛋白，大小相似。一个冷冻保护剂添加剂（CPA），冷却和变暖速率以及CPA稀释方法的范围将是测试。我们的目标是至少50％的生存率。 4：开发用于实现生存能力的测定法和效力。将测试4种方法：a）膜完整性，b）运动性，c）对蚊子的感染性产生PFSPZ，d）将软管转化为3天和8天的卵囊和IPFSPZ。