SBIR Phase I: Novel Urine-excreted Plasmodium Falciparum and P. vivax Poly-asparagine Proteins as Targets for Bi-specific Monoclonal Antibody and Urine Malaria Test Development

这个小型企业创新研究第一阶段项目旨在利用发热性疟疾患者尿液中的新型恶性疟原虫(PF)和间日疟原虫(PV)蛋白开发一种双价单抗。使用生物信息学方法，我们将在电子产生的自然抗原变体的重组体中创建、评分和选择，这些重组体旨在表达嵌合体，最大限度地将共同潜力包含在一个结构中，并将稀有表位的包含降至最低。重组抗原的抗原性通过使用来自表面健康的个体和临床肺原性疟疾和小儿麻痹症患者的一组特征良好的血清的间接ELISA法进行验证。使用来自PF/PV流行区的一系列临床尿样进行试纸试验，评估重组抗体的实用性。我们的总体目标是开发一种简单的一步尿液试纸试验，用于临床肺原性疟疾和/或PV疟疾的同时但特异的诊断。该项目更广泛的影响/商业潜力涉及开发一种基础广泛的一步尿液测试，用于在发烧患者中诊断临床肺炎和/或细小病毒疟疾。这项测试可能会提供一种可靠的工具来区分有症状和无症状的疾病，并为有需要的人提供更好的治疗选择。它有可能在疟疾流行地区，特别是在偏远地区和混合感染频繁发生的地方，通过消除静脉采血所涉及的职业危害，使临床疟疾诊断得到更广泛的接受，从而有助于减少不必要的抗疟疾药物的使用。总体而言，它有可能显著影响在疟疾流行地区诊断和治疗95%以上的所有临床疟疾的方式，并推动目前的全球努力，即在治疗所有发烧病例之前，按照世界卫生组织的要求对疟疾进行居家或医疗点检测。由于它基于与当前RDTs相同的平台，这种测试可以很容易地整合到当前的医疗结构中，为大多数流行国家的公共卫生提供重大好处。目前还没有这样的测试。

This Small Business Innovation Research Phase I project aims to develop a bivalent monoclonal antibody using novel Plasmodium falciparum (Pf) and P. vivax (Pv) proteins shed in the urine of febrile malaria patients. Using a bioinformatics approach, we will create, score, and select in silico-generated recombinants of natural antigen variants designed to express mosaics that maximize the inclusion of common potential in one structure and minimize the inclusion of rare epitopes. Antigenic properties of the recombinant antigens are validated by indirect ELISA using panels of well-characterized sera from apparently healthy individuals and clinical Pf and Pv malaria patients. The utility of the recombinant antibodies are evaluated in a dipstick test using a range of clinical urine samples from Pf/Pv endemic regions. Our overall objective is to develop a simple one-step urine-based dipstick test for the simultaneous but specific diagnosis of clinical Pf and/or Pv malaria. The broader impact/commercial potential of this project relates to the development of a broad-based one step urine test for home-based or point-of-care diagnosis of clinical Pf and/or Pv malaria in persons with fever. The test will potentially provide a reliable tool to differentiate between symptomatic and asymptomatic disease, and provide better treatment options to those in need. It will potentially offer wider acceptability of clinical malaria diagnosis in endemic regions, especially in remote settings and in places where mixed infections frequently occur by obviating the occupational hazards involved with phlebotomy, thus helping to reduce the unnecessary use of antimalarial drugs. Overall, it has the potential to markedly impact the way over 95% of all clinical malaria is diagnosed and treated in endemic regions, and drive the current global effort towards home-based or point-of-care testing for malaria mandated by the World Health Organization prior to treatment in all cases of fever. Since it is based on the same platform as current RDTs, this test can be easily integrated into current healthcare structures to provide significant benefits to public health in most endemic countries. No such test is currently available.