Human monoclonal antibodies for prophylaxis and therapy against the new coronavirus

用于预防和治疗新型冠状病毒的人单克隆抗体

基本信息

批准号：
10265634
负责人：
Michel C Nussenzweig
金额：
$ 45.83万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10265634
关键词：
2019-nCoV Advanced Development Antibodies B-Lymphocytes Binding Biological Assay Blood Blood Donations COVID-19 vaccine Cell Separation Cells China Chiroptera Cloning Collaborations Complex Coronavirus Coronavirus spike protein Cryoelectron Microscopy Crystallization Development Diagnostic Disease Disease Outbreaks Ebola Enzyme-Linked Immunosorbent Assay Epitopes Flavivirus Future Goals HIV HIV-1 Health Hepatitis B Virus Human Immunoglobulin G In Vitro Individual Infection Influenza Knowledge Laboratories Letters Light Malaria Medical Membrane Proteins Memory B-Lymphocyte Methods Middle East Respiratory Syndrome Coronavirus Molecular Molecular Conformation Mutation North Carolina Passive Transfer of Immunity Patients Pre-Clinical Model Procedures Prophylactic treatment Protein Binding Domain Proteins Protomer Recombinants Resistance SARS-CoV-2 spike protein Sampling Series Serum Severe Acute Respiratory Syndrome Structure Surface Antigens Techniques Testing Therapeutic Transfection Universities Ursidae Family Vaccine Design Vaccine Therapy Vaccines Variant Virus Washington ZIKA authority coronavirus disease cross reactivity effective therapy experimental study expression vector human monoclonal antibodies in vitro activity in vivo medical countermeasure neutralizing antibody novel coronavirus particle pathogen prevent protein complex protein structure public health emergency receptor binding recruit vaccine development

项目摘要

The WHO and US authorities have declared a public health emergency over the recent outbreak of a new coronavirus (CoV) originating from Wuhan, China (nCoV-2019, recently renamed SARS-CoV-2 and responsible for causing the coronavirus disease termed COVID-2019). The discovery of human monoclonal antibodies to this new CoV and obtaining a molecular understanding of its target epitopes will advance the development of diagnostics, therapeutics and vaccines to limit virus spread. The overall goal of this proposal is to discover and characterize potent broadly neutralizing antibodies to nCoV-2019 that also neutralize closely related strains of CoV such as SARS and other variants currently found in bats but likely to be able to produce human infections in the future. The Nussenzweig laboratory has developed robust methods to isolate, recombinantly produce and characterize human antibodies from the memory B cells of individuals infected by a series of different pathogens including HIV-1, Flaviviruses including Zika, and Hepatitis B virus (1, 2). These methods have also been used by other laboratories to isolate neutralizing antibodies to malaria, Ebola, influenza and other human infections (reviewed in (3)). The Bjorkman laboratory has performed structural studies using these antibodies to obtain information that directs vaccine design and therapies (2, 4-23). In Aim 1, we obtain samples from nCoV-2019 convalescing individuals (see letter from Dr. Wesley Van Voorhis). Serum samples will be tested for binding to the trimeric nCoV-2019 spike protein (S) and to the isolated receptor binding domain (RBD) of the S protein (see letter of collaboration from Dr. John Pak at Chan- Zuckerberg Biohub). Individuals with high titers against S and RBD will be recruited for large blood donations. Antibodies will be identified from the memory B cells of these individuals. In Aim 2 we will clone and express the antibodies obtained in Aim 1. The anti-nCoV-2019 antibodies will be tested for binding to the S protein from Severe Acute Respiratory Syndrome (SARS) and other closely related bat-derived CoV to test for cross- reactivity. Any promising antibodies will be evaluated for neutralizing activity (see letter by Dr. Timothy Sheahan at the University of North Carolina). In Aim 3 Dr. Bjorkman will solve crystal structures of antibody Fabs, and cryo-EM structures of coronavirus spike trimers complexed with Fabs from antibodies identified from Aims 1 and 2. In addition to helping guide vaccine development through the identification of neutralizing targets, the proposed discovery of human monoclonal antibodies to nCoV-2019 and related viruses bears a significant translational potential, such as the treatment and prophylaxis of severe medical conditions associated with nCoV-2019 infection by passive antibody transfer.

世界卫生组织和美国当局就最近爆发的新冠肺炎疫情宣布进入公共卫生紧急状态源自中国武汉的冠状病毒 (CoV)（nCoV-2019，最近更名为 SARS-CoV-2 和导致冠状病毒疾病（称为 COVID-2019）。人类单克隆抗体的发现针对这种新冠状病毒的抗体并获得对其目标表位的分子理解将推进开发诊断方法、治疗方法和疫苗以限制病毒传播。该提案的总体目标是发现并表征有效的广泛中和抗体 nCoV-2019 还可以中和与 CoV 密切相关的毒株，例如 SARS 和目前的其他变种在蝙蝠中发现了这种病毒，但将来可能会引起人类感染。努森茨威格实验室已开发出强大的方法来分离、重组生产和表征人类抗体被一系列不同病原体（包括 HIV-1、黄病毒）感染的个体的记忆 B 细胞包括寨卡病毒和乙型肝炎病毒 (1, 2)。其他实验室也使用这些方法分离出针对疟疾、埃博拉、流感和其他人类感染的中和抗体（在（3）中进行了综述）。这 Bjorkman 实验室使用这些抗体进行了结构研究，以获得指导的信息疫苗设计和疗法 (2, 4-23)。在目标 1 中，我们从 nCoV-2019 康复者中获取样本（参见 Wesley 博士的信）范·沃里斯）。将测试血清样本与三聚体 nCoV-2019 刺突蛋白 (S) 和 S 蛋白的分离受体结合域 (RBD)（参见 Chan-Pak 博士的合作信）扎克伯格生物中心）。抗S和RBD滴度高的个体将被招募进行大量献血。将从这些个体的记忆 B 细胞中鉴定出抗体。在目标 2 中，我们将克隆并表达目标 1 中获得的抗体。将测试抗 nCoV-2019 抗体与 S 蛋白的结合从严重急性呼吸系统综合症（SARS）和其他密切相关的蝙蝠衍生冠状病毒中进行交叉测试反应性。任何有希望的抗体都将进行中和活性评估（参见 Timothy 博士的信） Sheahan（北卡罗来纳大学）。在目标 3 中，Bjorkman 博士将解析抗体的晶体结构冠状病毒刺突三聚体的 Fab 和冷冻电镜结构与已鉴定的抗体中的 Fab 复合来自目标 1 和 2。除了通过识别中和靶点来帮助指导疫苗开发之外，提议发现针对 nCoV-2019 和相关病毒的人类单克隆抗体具有重要意义转化潜力，例如治疗和预防与以下疾病相关的严重医疗状况通过被动抗体转移感染 nCoV-2019。