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（由申请人提供）：该项目将完成多谱系床旁拉沙热诊断商业化的开发工作。拉沙热（LF）是一种严重的，往往致命的病毒性出血热（VHF）。由于其高病死率，易于通过人与人接触传播的能力，以及气溶胶释放的可能性，拉沙病毒（LASV），拉沙热的病原体，被列为生物安全4级和NIAID生物防御A类试剂。我们的团队已成功生产、验证、CE认证和商业化重组LASV床旁侧流免疫诊断（LFI），用于快速诊断由IV系病毒株引起的LF。这些检测是基于重组蛋白，而不是必须在高封闭实验室生产的试剂。我们还在塞拉利昂和尼日利亚建立了强有力的研究计划，这两个地区都是LASV的流行地区，为VHF研究提供了独特的临床和实验室资源。针对塞拉利昂和周边国家流行的LASV毒株开发的重组免疫测定法现在将重新配置用于尼日利亚的三种不同的LASV谱系--因为谱系IV特异性RDT对由毒株变异引起的谱系I-III的循环病毒缺乏敏感性。我们现在将在后期开发中执行关键步骤，以实现多谱系护理点LF诊断的商业化。在MILESTONE 1中，我们将使用充分表征的血清开发组完成I-IV系LASV的商业级LASV抗原捕获和免疫球蛋白M（IgM）和免疫球蛋白G（IgG）抗体捕获酶联免疫吸附试验（ELISA）的开发。在MILESTONE 2中，我们将使用开发血清组合和新开发的抗体完成LASV多谱系LFI的开发，作为床旁诊断。在MILESTONE 3中，我们将根据良好生产程序（GMP）生产多谱系重组ELISA和LFI，以提供足够的商业级诊断试剂盒，用于设计控制参数的临床前评价， 临床研究、CE认证和商业化所需的基准。在MILESTONE 4中，我们将优化代表当前循环的LASV谱系I-IV的重组LASV核蛋白（NP）的放大和纯化，以及识别所有当前循环的LASV谱系I-IV的重组LASV NP抗体的放大/纯化方法。然后，我们将转移到生产，以提供足以用于商业检测开发和生产的重组蛋白和抗体。在MILESTONE 5中，我们将从西非和其他地方的LASV流行范围内的不同地区（欧洲和美国对照）确定并收集阳性和阴性血清用于检测验证。然后，我们将验证多谱系LF重组ELISA和LFI的灵敏度和特异性。在MILESTONE 6中，我们将编制多谱系reLASV RDT的设计确认报告，提交给欧盟委员会进行CE认证，并提交给NAFDAC进行尼日利亚的产品注册。