Preclinical development of recombinant antigen diagnostics for Lassa fever

拉沙热重组抗原诊断的临床前开发

• 批准号: 8458569
• 负责人: Robert F Garry
• 金额: $ 123.72万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-22 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8458569
• 关键词: Achievement; Aerosols; Africa; Antibodies; Antigens; Area; Benchmarking; Biological; Biological Assay; Case Fatality Rates; Categories; Clinical; Containment; Development; Diagnosis; Diagnostic; Enzyme-Linked Immunosorbent Assay; Health Benefit; Human; Immunoglobulin G; Immunoglobulin M; Individual; Instruction; Laboratories; Lassa Fever; Lassa virus; Lateral; Medical; Military Personnel; Monoclonal Antibodies; National Institute of Allergy and Infectious Disease; Performance; Process; Production; Public Health; Quality Control; Reagent; Recombinant Proteins; Recombinants; Research; Resources; Serum; Sierra Leone; Testing; Validation; Viral Antigens; Viral Hemorrhagic Fevers; Viral Proteins; base; biodefense; cost effective; design; hospital laboratories; method development; point of care; point-of-care diagnostics; pre-clinical; preclinical evaluation; programs; prototype; quality assurance; scale up; weapons

DESCRIPTION (provided by applicant): This project will develop Diagnostics For Biodefense against Lassa fever, a severe, often fatal viral hemorrhagic fever (VHF). Because of its high case fatality rate, ability to 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 prototype LASV enzyme-linked immunosorbent assays (ELISA) that are based on recombinant proteins rather than on reagents that must be produced in high containment laboratories. We have also newly established a research program in Sierra Leone, an area endemic for LASV, that provides unique clinical and laboratory resources for VHF research. We will now perform critical steps in the preclinical development of commercial recombinant antigen Lassa fever diagnostics. In MILESTONE 1 we will development commercial Lassa fever recombinant antigen-capture and immunoglobulin M (IgM) and immunoglobulin G (IgG) antibody-capture ELISA using a Performance Panel of well-characterized sera. In MILESTONE 2 we will development Lassa fever recombinant lateral flow point-of-care diagnostics. In MILESTONE 3 we will convert to manufacturing recombinant ELISA and lateral flow assays under Good Manufacturing Practices (GMP) with Quality Assurance (QA)/Quality Control (QC) to provide quantities of commercial grade diagnostic kits sufficient for preclinical evaluation of design control parameters to achieve benchmarks required for FDA approval. In MILESTONE 4 we will optimize scale up/purification of recombinant LASV proteins, GP1, GP2, and NP and monoclonal antibodies (mAbs) to recombinant LASV GP1, GP2, and NP. We will also convert to manufacturing with QA/QC, to provide quantities of recombinant proteins and mAbs sufficient for development and testing of commercial assays. In MILESTONE 5 we will define and collect positive and negative sera for assay validation from diverse regions across the Lassa fever endemic range of West Africa and elsewhere. These sera will be used to test and compare the commercial LASV recombinant ELISA and recombinant lateral flow point-of-care diagnostic assays with results from BSL-4 ELISA and PCR assays. RELEVANCE (See instructions): The potential use of LASV as a biological weapon necessitates development of methods to diagnose individuals exposed to and/or infected with LASV. The impact of Lassa fever in endemic areas of West Africa is immense, and a safe and effective diagnostic can also provide a very significant public health benefit.

描述（由申请人提供）：该项目将开发针对拉沙热的生物防御诊断，拉沙热是一种严重且往往致命的病毒性出血热 (VHF)。拉沙热的病原体拉沙病毒 (LASV) 由于其病死率高、通过人与人接触容易传播的能力以及气溶胶释放的可能性，被列为生物安全 4 级和 NIAID 生物防御 A 类病原体。我们的团队已成功生产出 LASV 酶联免疫吸附测定 (ELISA) 原型，该测定基于重组蛋白，而不是基于必须在高安全性实验室中生产的试剂。我们还在 LASV 流行地区塞拉利昂新建立了一个研究项目，为 VHF 研究提供独特的临床和实验室资源。我们现在将在商业重组抗原拉沙热诊断的临床前开发中执行关键步骤。在 MILESTONE 1 中，我们将使用经过充分表征的血清性能面板来开发商业拉沙热重组抗原捕获以及免疫球蛋白 M (IgM) 和免疫球蛋白 G (IgG) 抗体捕获 ELISA。在 MILESTONE 2 中，我们将开发拉沙热重组侧流护理点诊断。在 MILESTONE 3 中，我们将转而按照良好生产规范 (GMP) 和质量保证 (QA)/质量控制 (QC) 生产重组 ELISA 和侧向层析检测，以提供足够数量的商业级诊断试剂盒，用于设计控制参数的临床前评估达到 FDA 批准所需的基准。在 MILESTONE 4 中，我们将优化重组 LASV 蛋白、GP1、GP2 和 NP 以及重组 LASV GP1、GP2 和 NP 的单克隆抗体 (mAb) 的放大/纯化。我们还将转向通过 QA/QC 进行生产，以提供足够数量的重组蛋白和单克隆抗体，用于商业化验的开发和测试。在里程碑 5 中，我们将定义并收集西非和其他拉沙热流行区不同地区的阳性和阴性血清，以进行检测验证。这些血清将用于测试商业 LASV 重组 ELISA 和重组侧流护理点诊断测定，并将其与 BSL-4 ELISA 和 PCR 测定的结果进行比较。相关性（参见说明）：LASV 作为生物武器的潜在用途需要开发方法来诊断暴露于和/或感染 LASV 的个体。拉沙热在西非流行地区的影响是巨大的，安全有效的诊断也可以提供非常显着的公共卫生益处。

项目成果

Inhibition of arenavirus infection by a glycoprotein-derived peptide with a novel mechanism.

糖蛋白衍生肽以新机制抑制沙粒病毒感染。

• DOI: 10.1128/jvi.01133-14
• 发表时间: 2014
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Spence,JenniferS;Melnik,LiliaI;Badani,Hussain;Wimley,WilliamC;Garry,RobertF
• 通讯作者: Garry,RobertF

Human-monoclonal-antibody therapy protects nonhuman primates against advanced Lassa fever.

• DOI: 10.1038/nm.4396
• 发表时间: 2017-10
• 期刊: Nature medicine
• 影响因子: 82.9
• 作者: Mire CE;Cross RW;Geisbert JB;Borisevich V;Agans KN;Deer DJ;Heinrich ML;Rowland MM;Goba A;Momoh M;Boisen ML;Grant DS;Fullah M;Khan SH;Fenton KA;Robinson JE;Branco LM;Garry RF;Geisbert TW
• 通讯作者: Geisbert TW

Lassa virus-like particles displaying all major immunological determinants as a vaccine candidate for Lassa hemorrhagic fever.

• DOI: 10.1186/1743-422x-7-279
• 发表时间: 2010-10-20
• 期刊: Virology journal
• 影响因子: 4.8
• 作者: Branco LM;Grove JN;Geske FJ;Boisen ML;Muncy IJ;Magliato SA;Henderson LA;Schoepp RJ;Cashman KA;Hensley LE;Garry RF
• 通讯作者: Garry RF

Treatment of Lassa virus infection in outbred guinea pigs with first-in-class human monoclonal antibodies.

• DOI: 10.1016/j.antiviral.2016.08.012
• 发表时间: 2016-09
• 期刊: ANTIVIRAL RESEARCH
• 影响因子: 7.6
• 作者: Cross, Robert W.;Mire, Chad E.;Branco, Luis M.;Geisbert, Joan B.;Rowland, Megan M.;Heinrich, Megan L.;Goba, Augustine;Momoh, Mambu;Grant, Donald S.;Fullah, Mohamed;Khan, Sheik Humarr;Robinson, James E.;Geisbert, Thomas W.;Garry, Robert F.
• 通讯作者: Garry, Robert F.