Late-stage development toward commercialization of multilineage point-of-care Lassa fever diagnostics

多谱系护理点拉沙热诊断商业化的后期开发

• 批准号: 9003026
• 负责人: Luis Manuel Branco
• 金额: $ 92.85万
• 依托单位: ZALGEN LABS, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-05 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9003026
• 关键词: Achievement; Acute; Aerosols; Africa; Antibodies; Antigens; Area; Benchmarking; Biological; Biological Assay; Biological Warfare; Caring; Case Fatality Rates; Categories; Central Africa; Clinical; Clinical Research; Containment; Country; Developed Countries; Development; Diagnosis; Diagnostic; Enzyme-Linked Immunosorbent Assay; European; Guinea; Health; Health Benefit; Human; Immunoassay; Immunoglobulin G; Immunoglobulin M; Immunological Diagnosis; Industry; Investments; Laboratories; Lassa Fever; Lassa virus; Lateral; Liberia; Medical; Methods; Military Personnel; Monoclonal Antibodies; National Institute of Allergy and Infectious Disease; Nigeria; Nucleoproteins; Procedures; Process; Production; Public Health; Public Hospitals; Reagent; Recombinant Antibody; Recombinant Proteins; Recombinants; Reporting; Research; Resources; Sensitivity and Specificity; Serum; Sierra Leone; Staging; Technology; Testing; Validation; Variant; Viral Antigens; Viral Hemorrhagic Fevers; Virus; Virus Diseases; base; biodefense; biothreat; commercialization; cost effective; design; field study; point of care; point-of-care diagnostics; polyclonal antibody; preclinical evaluation; programs; prototype; rapid diagnosis; scale up; weapons

 DESCRIPTION (provided by applicant): This project will complete development efforts toward commercialization of multilineage point-of-care Lassa fever diagnostics. Lassa fever (LF) is a severe, often fatal viral hemorrhagic fever (VHF). Because of its high case fatality rate, ability o spread easily by human-human contact, and potential for aerosol release, Lassa virus (LASV), the causative agent of Lassa fever, is classified as a Biosafety Level 4 and NIAID Biodefense category A agent. Our team has successfully produced, validated, CE marked and commercialized recombinant LASV point-of-care lateral flow immunodiagnostics (LFI) for rapid diagnosis of LF caused by lineage IV virus strains. These assays are based on recombinant proteins rather than on reagents that must be produced in high containment laboratories. We have also established robust research programs in Sierra Leone and Nigeria, both endemic areas for LASV, that provide unique clinical and laboratory resources for VHF research. The recombinant immunoassays developed for strains of LASV prevalent in Sierra Leone and surrounding countries will now be reconfigured for the three divergent lineages of LASV in Nigeria - due to lack of sensitivity of lineage IV-specific RDTs for circulating viruses from lineages I-III arising from strain variation. We will now perform critical steps in late-stage development toward commercialization of multilineage point-of- care LF diagnostics. In MILESTONE 1 we will complete development of commercial grade LASV antigen- capture and immunoglobulin M (IgM) and immunoglobulin G (IgG) antibody-capture enzyme-linked immunosorbent assays (ELISA) to lineage I-IV LASV using a Developmental Panel of well-characterized sera. In MILESTONE 2 we will complete development of LASV multilineage LFI as point-of-care diagnostics using the Developmental Serum Panel and newly developed antibodies. In MILESTONE 3 we will convert to manufacturing multilineage recombinant ELISA and LFI under Good Manufacturing Procedures (GMP) to provide quantities of commercial grade diagnostic kits sufficient for preclinical evaluation of design control parameters to achieve benchmarks required for clinical studies, CE marking, and commercialization. In MILESTONE 4 we will optimize scale up and purification of recombinant LASV nucleoprotein (NP) representing currently circulating LASV lineages I-IV, and scale up/purification methods for antibodies to recombinant LASV NP recognizing all currently circulating LASV lineages I-IV. We will then transfer to manufacturing to provide quantities of recombinant proteins and antibodies sufficient for development and production of commercial assays. In MILESTONE 5 we will define and collect positive and negative sera for assay validation from diverse regions across the LASV endemic range of West Africa and elsewhere (European and U.S. controls). We will then validate sensitivity and specificity of multilineage LF recombinant ELISA and LFI. In MILESTONE 6 we will compile Design Validation Report for multilineage reLASV RDT, submission to European Commission for CE marking and to NAFDAC for product registration in Nigeria.

 描述（由应用程序提供）：该项目将完成开发努力，用于商业化多收入级别的LASSA发烧诊断。 LASSA热（LF）是一种严重的，通常是致命的病毒出血热（VHF）。由于其较高的病例死亡率，能力o很容易通过人类接触而传播，并且释放气溶胶的潜力，LASSA病毒（LASV）是LASSA发烧的致病药物，被归类为生物安全水平4和NIAID Biodefense A类药物。我们的团队已成功生产，经过验证，CE标记和商业化重组LASV护理点侧流量免疫诊断（LFI），用于谱系IV病毒菌株引起的LF的快速诊断。这些测定基于重组蛋白，而不是基于必须在高遏制实验室中生产的试剂。我们还在塞拉利昂（Sierra Leone）和尼日利亚（Nigeria）建立了强大的研究计划，这都是LASV的内在区域，为VHF研究提供了独特的临床和实验室资源。现在，塞拉利昂及周边国家 /周边国家为LASV菌株开发的重组免疫测定现在将重新配置尼日利亚LASV的三种不同谱系 - 由于缺乏因应变的I-i-i-iiii raviriation raviriation raviriation cariation riatiation cartiriation carliation cartiriation frol lineage rdts的谱系IV特异性RDT的敏感性。现在，我们将在晚期开发中执行关键步骤，以实现多列纳格LF诊断的商业化。在里程碑1中，我们将完成商业级LASV抗原捕获和免疫球蛋白M（IGM）和免疫球蛋白G（IGG）抗体捕获酶链接的免疫吸收测定法（ELISA），用于使用良好的Sera的开发面板。在Milestone 2中，我们将使用发育血清面板和新开发的抗体来完成LASV Multineage LFI作为护理诊断的开发。在里程碑3中，我们将在良好的制造程序（GMP）下转换为制造的多列元素重组ELISA和LFI，以提供数量的商业级诊断套件，足以对设计控制参数进行临床前评估以实现 临床研究，CE标记和商业化所需的基准。在里程碑4中，我们将优化代表当前循环LASV谱系I-IV的重组LASV核蛋白（NP）的规模和纯化，并对重组LASV NP的抗体进行扩展/纯化方法，以识别所有当前循环LASV LASV LASV LASV LASV LASV LASEAGE lASV LASV IV。然后，我们将转移到制造中，以提供足以开发和生产商业测定的重组蛋白和抗体。在Milestone 5中，我们将定义并收集来自西非和其他地方LASV内在范围内的潜水区域的测定验证的正阳性血清（欧洲和美国对照组）。然后，我们将验证Multineage LF重组ELISA和LFI的灵敏度和特异性。在Milestone 6中，我们将编译有关Multilineage Relasv RDT的设计验证报告，提交给欧洲CE Marking委员会和NAFDAC的尼日利亚NAFDAC。