Cryopreservation of Anopheles Embryos

按蚊胚胎的冷冻保存

• 批准号: 10377924
• 负责人: Peter F. Billingsley
• 金额: $ 96.68万
• 依托单位: SANARIA, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377924
• 关键词: Adult; Africa; Africa South of the Sahara; Anopheles Genus; Attenuated; Biological Assay; Biological Products; Blood; Breeding; Burkina Faso; Businesses; Certification; Child; Clinical Trials; Communities; Conduct Clinical Trials; Cryopreservation; Culicidae; Cyclic GMP; Data; Development; Dose; Embryo; Ensure; Equatorial Guinea; Equipment Failure; Event; Exposure to; Falciparum Malaria; Female; Funding; Future; Gabon; Generations; Genetic Drift; Geographic Locations; Germany; Goals; Harvest; Human; Infection; Laboratories; Laboratory Research; Larva; Licensing; Life Cycle Stages; Malaria; Malaria Vaccines; Mali; Metabolic; Methodology; Methods; Molecular Genetics; Mosquito Control; Natural Immunity; Natural Resistance; Parasitic Diseases; Partner in relationship; Persons; Phase; Phase III Clinical Trials; Phenotype; Plasmodium falciparum; Plasmodium falciparum vaccine; Population; Preparation; Process; Production; Publications; Quality Control; Radiation; Readiness; Regimen; Research; Research Personnel; Resources; Resuscitation; Salivary Glands; Services; Small Business Innovation Research Grant; Source; Sporozoites; Tanzania; Technology; Testing; Time; Vaccines; Vial device; Visit; Work; arm; attenuation; base; cold temperature; cryogenics; drug candidate; egg; gamma irradiation; malaria infection; malaria transmission; manufacturing process; novel; oocyte cryopreservation; programs; public health relevance; research clinical testing; scale up; success; tool; vaccine candidate; vaccine efficacy; zygote

ABSTRACT: PfSPZ Vaccine is currently on track to be the world’s first licensed vaccine against malaria and the first vaccine against a human parasitic disease. PfSPZ Vaccine is composed of aseptic, attenuated, metabolically active, purified, vialed and cryopreserved Plasmodium falciparum (Pf) sporozoites (SPZ). Manufacture of PfSPZ Vaccine shares all steps (except radiation attenuation) with PfSPZ Challenge that is used by researchers worldwide in controlled human malaria infections to evaluate experimental malaria drugs, candidate vaccines, and natural immunity or resistance to malaria. The third product under clinical testing is PfSPZ-CVac, PfSPZ Challenge given under chemoprophylactic cover. PfSPZ Vaccine has protected against homologous (same strain) and heterologous (different strains) of Plasmodium falciparum (Pf) malaria in clinical trials conducted in the U.S., Germany, Tanzania, and Mali. To date, 80–100% protection lasting for at least 8 months against heterologous and 14 months against homologous challenge infections has been demonstrated. Ongoing trials in Gabon, Mali, and Equatorial Guinea are testing optimized regimens in preparation for Phase 3 clinical trials starting in 2020 in support of a Biologics License Application (BLA) to the FDA in 2021. All PfSPZ products are produced in mosquitoes reared aseptically from disinfected eggs that are harvested from a continuously-maintained Anopheles stephensi colony. The goals of this project are to submit a Biologics Master File for Sanaria’s cryopreserved A. stephensi SDA500 strain in support of the manufacture of PfSPZ- based products and to manufacture one lot of PfSPZ Challenge using mosquitoes derived from cryopreserved eggs, and also to adapt the methodology for large scale cryopreservation of A. gambiae. Fundamental to this process is the elevation of mosquito production from banks of cryopreserved eggs at full cGMP compliance. The work will involve cryopreservation of eggs at hugely increased scale and the generation of a master egg bank (MEB) and working egg bank (WEB) of cryopreserved A. stephensi SDA500 eggs. The work is incorporated into three specific aims: 1) Complete the GMP-compliant manufacture of a MEB and WEB which will entail development of novel release and stability assays, generation of SOPs and Certificates of Analysis, establishment of a stability program for cryopreserved eggs and submission of a Biologics Master File to the FDA; 2) Manufacture, using mosquitoes derived from the cryopreserved WEB, one lot of PfSPZ Challenge for use in clinical trials; and 3) Cryopreserve A. gambiae eggs at scale to support mosquito release programs. Cryopreservation of Anopheles eggs is a breakthrough enabling technology that will be available to support Anopheles mosquito research, particularly in laboratories that need to retain genetically modified strains and lines but have limited resources to do so.

摘要：PfSPZ 疫苗目前有望成为世界上第一个获得许可的针对疟疾和疟疾的疫苗。 第一种针对人类寄生虫病的疫苗。 PfSPZ 疫苗由无菌、减毒、 代谢活跃、纯化、装瓶并冷冻保存的恶性疟原虫 (Pf) 子孢子 (SPZ)。 PfSPZ 疫苗的制造与 PfSPZ Challenge 共享所有步骤（辐射衰减除外），即 世界各地的研究人员使用受控人类疟疾感染来评估实验性疟疾药物， 候选疫苗以及对疟疾的自然免疫力或抵抗力。第三个进入临床试验的产品是 PfSPZ-CVac、PfSPZ 挑战在化学预防保险范围内进行。 PfSPZ 疫苗可预防 临床上恶性疟原虫（Pf）疟疾的同源（同一菌株）和异源（不同菌株） 试验在美国、德国、坦桑尼亚和马里进行。迄今为止，80–100% 的保护持续至少 8 已证明可抵抗异源感染 14 个月，抵抗同源攻击感染 14 个月。 加蓬、马里和赤道几内亚正在进行的试验正在测试优化的治疗方案，为第三阶段做准备 临床试验将于 2020 年开始，以支持 2021 年向 FDA 提交的生物制剂许可申请 (BLA)。所有 PfSPZ 产品是在无菌饲养的蚊子中生产的，这些蚊子是从经过消毒的卵中收获的。 持续维持的史氏按蚊群体。该项目的目标是提交一份生物制剂 Sanaria 冷冻保存的 A. stephensi SDA500 菌株的主文件，用于支持 PfSPZ- 的生产 基于产品并使用冷冻保存的蚊子生产一批 PfSPZ Challenge 卵，并适应冈比亚 A. 大规模冷冻保存的方法。对此的根本 该过程是在完全符合 cGMP 的情况下提高冷冻卵库中的蚊子产量。 这项工作将涉及大幅增加规模的卵子冷冻保存以及主卵的产生 冷冻保存的 A. stephensi SDA500 卵的库 (MEB) 和工作卵库 (WEB)。工作是 纳入三个具体目标： 1) 完成符合 GMP 要求的 MEB 和 WEB 制造， 将需要开发新型释放和稳定性测定方法、生成标准操作程序和分析证书， 建立冷冻卵的稳定性计划并将生物制品主文件提交给 美国食品和药物管理局； 2) 使用来自冷冻保存的 WEB 的蚊子制造一批 PfSPZ Challenge 用于临床试验； 3) 大规模冷冻保存冈比亚蚊卵以支持蚊子释放计划。 按蚊卵的冷冻保存是一项突破性的技术，可用于支持 按蚊研究，特别是在需要保留转基因菌株和 但资源有限。

项目成果

Cryopreservation of Anopheles stephensi embryos.

• DOI: 10.1038/s41598-021-04113-x
• 发表时间: 2022-01-07
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: James ER;Wen Y;Overby J;Pluchino K;McTighe S;Matheny S;Eappen A;Hoffman SL;Billingsley PF
• 通讯作者: Billingsley PF