Structure-guided neutralizing antibodies developed using EpiVolve technology

摘要 目前开发中和抗体的策略并不有效，通常涉及从 康复的病人。大流行病毒进化为突变，可以保护其表位免受宿主免疫 监控系统，因此许多重要的表位将被遗漏。即使中和了抗体，分离出来的抗体也是 从人类样本中，它们还需要进一步利用表位结合和测定 避免脱靶效应的特殊性。探索病毒整个蛋白质表面的系统方法 能够识别病毒上所有可能影响其生命周期的潜在位置将具有重大影响 是必要的。我们提出了一种结构导向的系统抗体开发流水线来发现抗体 可以对抗传染病。我们建议使用我们的新型定点抗体开发技术， EpiVolve(表位进化的缩写)。EpiVolve将用于开发暴露于溶剂中的部位特异性抗体 残基和相邻的‘上下文’序列。这些抗体将用于对抗传染病。这个 EpiVolve的优点是：a)克服免疫耐受，靶向病毒的模仿人类蛋白质组 表位，b)精确地针对任何抗原表位，而不考虑其免疫原性，c)利用 B细胞扩增和体细胞超突变以产生针对一个靶向残基的IgG克隆型， 其允许生成泛变异体抗体和特定多态抗体两者的能力，以及d)能力 用于产生中和抗体发现管道。我们将以SARS-Cov-2病毒为模型，在第一阶段和 第二阶段甲型流感。EpiVolve开发的部位特异性抗体将针对暴露在溶剂中的残留物 蛋白质表面。结构导向的银设计将使EpiVolve技术在这一系统中得到支持 分析。就这项建议，我们会提供有关沙士的EpiVolve试验研究的最新初步数据： CoV-2受体结合域(RBD)，主要集中在宿主细胞受体ACE2结合界面。为 第一阶段研究，我们建议完成初步研究，并将研究扩展至整个蛋白质表面。 RBD域。通过结合亲和力和中和SARS-CoV-2病毒的能力来表征每一种抗体 将被纳入第一阶段研究。在第二阶段，我们建议把第一阶段研究所得的经验应用于 另一种非常重要的病毒模型是甲型流感病毒。特别针对溶剂可及的 血凝素(HA)蛋白的茎/柄保守区残基

Abstract Current strategies for developing neutralizing Abs are not effective and typically involve screening IgGs from recovered patients. Pandemic viruses evolve for mutations that can shield their epitopes from host immune surveillance system, so a lot of important epitopes will be missed. Even after neutralizing Abs isolated are from human sample, they still need further characterization using epitope binning and determination of specificities to avoid off target effect. A systematic method for exploring the entire protein surface of a virus that can identify all potential sites on the virus which can affect its life cycle would have significant impact and is needed. We propose a structure-guided systematic Ab development pipeline to discover Abs that can fight infectious diseases. We propose using our novel site-directed Ab development technology, ‘EpiVolve’ (short for Epitope Evolution). EpiVolve will be used to develop site-specific Abs to solvent-exposed residues and the adjacent ‘context’ sequences. These Abs will be used for fighting infectious disease. The advantages of EpiVolve are a) overcoming immune tolerance and targeting virus’ human proteome-mimicking epitopes, b) precisely targeting any antigenic epitopes regardless of its immunogenicity, c) taking advantage of B cell expansion and somatic hypermutation to generate IgG clonotypes against one targeted residue, which allows an ability to generate both pan-variants Abs and polymorphism-specific Abs, and d) an ability for generating a neutralizing Ab discovery pipeline. We will model this on SARS-cov-2 virus in Phase I and Influenza A in Phase II. EpiVolve developed site-specific antibodies will target solvent-exposed residues on the protein surface. Structure-guided Ag design will empower the EpiVolve technology in this systematic analysis. For this proposal, we will present the current preliminary data on the pilot EpiVolve study on SARS- cov-2 Receptor Binding Domain (RBD), focused mainly on the host cell receptor ACE2 binding interface. For Phase I studies, we propose to complete the pilot study and extend the study to the whole protein surface of the RBD domain. Characterizing each Ab by its binding affinity and ability to neutralize SARS-cov-2 virus will be included in Phase I studies. For Phase II, we propose to apply the learnings from this Phase I study on another virus model of great importance, the Influenza A virus. Specifically targeting the solvent-accessible residues of the conserved Stem/Stalk region of the Hemagglutinin (HA) protein