Automating mosquito microdissection for a malaria PfSPZ vaccine

描述：疟疾每年导致2.5亿临床病例和约100万人死亡，造成非洲国内生产总值1%的损失，是旅行者和军事人员严重关切的问题。人们迫切需要一种高效的疫苗，经蚊子叮咬接种的辐射减毒恶性疟原虫(Pf)子孢子(SPZ)和含有氯喹化学预防的非减毒PfSPZ(用子孢子化学预防，CPS)对人类疟疾具有最有效(90%)和持久(10-28个月)的保护作用。在最近的临床试验中，Sanaria的PfSPZ疫苗，包括辐射减毒、无菌、纯化、冷冻保存的PfSPZ，保护了6/6(100%)的志愿者，6/9名受试者静脉注射(IV)最高剂量和第二高剂量的PfSPZ，这在疟疾疫苗发展史上是前所未有的。明年，PfSPZ疫苗将在针对美国、德国、坦桑尼亚、马里和赤道几内亚的个人的临床试验中进行严格的测试，以期在大约4年内获得在非洲开展大规模管理消除运动和在旅行者中预防疟疾的许可。此外，基于Sanaria的非减毒、传染性PfSPZ(PfSPZ挑战)可以感染欧洲和美国100%的志愿者的证明，PfSPZ挑战与化学预防相结合是Sanaria的第二种疫苗产品PfSPZ-化学预防(C)疫苗(Vac)(PfSPZ-CVac)。获得许可的加速之路需要扩大努力，以满足预计的市场需求。尽管每年所需的PfSPZ绝对数将取决于PfSPZ/剂量的数量，以及每只蚊子可以获得的PfSPZ的数量，但我们预计很快就需要每年处理10^7只蚊子，然后是10^8只蚊子，最后是2x10^8只蚊子。手动解剖10^7只蚊子/年是可以通过在我们当前的制造设施内扩大2倍来实现的，然而，自动化可以促进在相同的物理空间内进一步增加5到10倍的吞吐量(5x10^7-10^8只蚊子/年)。相比之下，每年手工解剖10^8只蚊子将需要相当大的扩张，才能容纳一个庞大的解剖工作组。对预计成本曲线的建模也表明疫苗的价格将会下降 只有在自动化到位，而不是手动解剖的情况下，才会逐步制造每一个增量。事实上，在处理了第一批10^7只蚊子后，疫苗的每瓶成本下降了50%。在初步开发可自动化原型的基础上，我们建议建立第一个集成的自动蚊子显微解剖系统(AMMS)，并微调性能参数和材料顺应性，以实现工业化规模的高通量cGMP加工。这些目标的成功完成将使Sanaria的制造进展与其临床开发和商业化计划步入正轨，使现有设施的产量大大提高，而不需要进行重大扩张，从而促进更快地为数亿患有这种疾病的人广泛使用完整的PfSPZ疫苗。

DESCRIPTION: Malaria causes >250 million clinical cases and ~1 million deaths annually, is responsible for >1% loss of GDP in Africa and is a serious concern for travelers and military personnel. A highly effective vaccine is desperately needed and radiation-attenuated Plasmodium falciparum (Pf) sporozoites (SPZ) and non- attenuated PfSPZ with chloroquine chemoprophylaxis (ChemoProphylaxis with Sporozoites, CPS), administered by mosquito bites, induce the most efficacious (>90%) and long lasting (10-28 months) protection against malaria in humans. In its most recent clinical trial, Sanaria's PfSPZ Vaccine, consisting of radiation attenuated, aseptic, purified, cryopreserved PfSPZ, protected 6/6 (100%) of volunteers 6/9 subjects who received the highest and second highest dose of PfSPZ intravenously (IV), unparalleled in the history of malaria vaccine development. PfSPZ Vaccine will undergo rigorous testing during the next year in clinical trials targeting individuals in the U.S., Germany, Tanzani, Mali, and Equatorial Guinea towards licensure in ~4 years for mass administration elimination campaigns in Africa and prevention of malaria in travelers. Also, based on the demonstration that Sanaria's non-attenuated, infectious PfSPZ (PfSPZ Challenge) can infect 100% of volunteers in Europe and the US, PfSPZ Challenge in conjunction with chemoprophylaxis is Sanaria's 2nd vaccine product, PfSPZ-Chemoprophylaxis (C) Vaccine (Vac) (PfSPZ-CVac). The accelerated path to licensure requires scale-up efforts to meet projected market needs. Although the absolute number of PfSPZ required/yr will depend on the numbers of PfSPZ/dose, and the numbers of PfSPZ that can be obtained per mosquito, we anticipate soon needing to process 10^7, then 10^8 and finally >2x10^8 mosquitoes/yr. Manual dissection of 10^7 mosquitoes/yr is achievable with a 2-fold expansion within our current manufacturing facility however, automation can facilitate a further 5 to 10-fold increase (5x10^7 -10^8 mosquitoes/yr) in throughput within the same physical space. In contrast dissection of 10^8 mosquitoes per year by hand will require sizeable expansion in order to house a hugely expanded dissector taskforce. Modeling of projected cost curves also indicates that the price of the vaccine will drop progressively with each increment manufactured only if automation is put in place, but not with manual-dissection. In fact after processing the first 10^7 mosquitoes, the cost/vial of vaccine drops >50%. Based on preliminary development of automatable prototypes we propose to construct the first integrated automated mosquito microdissection system (AMMS) and fine-tune performance parameters and materials compliance for high throughput cGMP processing at an industrial scale. Successful completion of these goals will put Sanaria's manufacturing progress on track with its clinical development and commercialization plans, allow for greatly enhanced production from current facilities without a need for significant expansion and thereby facilitate more immediate widespread use of a whole PfSPZ vaccine for the hundreds of millions who suffer from this disease.