Human monoclonal antibodies for prophylaxis and therapy against the new coronavirus

世界卫生组织和美国当局宣布最近爆发一种新的公共卫生紧急状态 起源于武汉的冠状病毒中国(NCoV-2019年，最近更名为SARS-CoV-2和 导致冠状病毒病的责任被称为COVID-2019)。人类单克隆体的发现 对这种新冠状病毒的抗体和对其目标表位的分子理解将推动 开发诊断、治疗和疫苗以限制病毒传播。 这项提议的总体目标是发现和表征有效的广谱中和抗体。 到nCoV-2019，也中和密切相关的冠状病毒毒株，如SARS和目前的其他变种 在蝙蝠身上发现，但将来很可能会引起人类感染。努森茨韦格实验室 已经开发出强大的方法来分离、重组生产和鉴定人类抗体 包括HIV-1、黄病毒在内的一系列不同病原体感染个体的记忆B细胞 包括寨卡病毒和乙肝病毒(1，2)。这些方法也被其他实验室用来 分离疟疾、埃博拉、流感和其他人类感染的中和抗体(在(3)中回顾)。这个 比约克曼实验室使用这些抗体进行了结构研究，以获得指导 疫苗设计和治疗(2、4-23)。 在目标1中，我们从nCoV-2019康复者身上获取样本(见Wesley博士的信 范·沃里斯)。将检测血清样本与NCoV-2019三聚体刺突蛋白(S)和 分离的S蛋白的受体结合域(见白约翰博士在Chan- 扎克伯格生物中心)。S和RBD抗体效价高的个人将被招募进行大量献血。 抗体将从这些人的记忆B细胞中识别出来。在目标2中，我们将克隆和表达 目的1.检测抗新冠病毒-2019年抗体与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到测试交叉 反应性。任何有希望的抗体都将被评估中和活性(见蒂莫西博士的信 北卡罗来纳大学的Sheahan)。在目标3中，比约克曼博士将解决抗体的晶体结构 冠状病毒刺突三聚体与抗体复合后的FABS和冷冻-EM结构 来自目标1和目标2。 除了通过识别中和目标来帮助指导疫苗开发外， 拟议中发现的针对nCoV-2019及相关病毒的人单抗具有重大意义 转化潜力，如治疗和预防与以下疾病相关的严重疾病 NCoV-2019通过被动抗体转移感染。

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.