Rapid antibody screening systems to identify and engineer antiviral protection

项目摘要 对分子和细胞适应性免疫反应的详细了解对于加速 人类免疫学和药物开发的进展。然而，用于分析抗病毒药物的现有技术 中和抗体应答对于大规模临床样品分析是缓慢且不切实际的，并且可以提供 关于人体免疫中中和抗体特征范围的信息有限。重要的是，当前 这些方法也不能工程化抗体分子以直接提高中和效力，这是一个缺点。 这是发现针对病毒性疾病的有效和广泛反应性的基于抗体的干预措施的主要限制。 目前的方法也不能工程化针对相关病毒的广泛抗体中和，这对于免疫缺陷病毒至关重要。 针对不同病毒谱系的药物和疫苗开发。重要的例子包括不同的病毒 β冠状病毒和黄病毒的谱系，其中针对进化和扩展的病毒谱系的保护是 对于有效的临床应用至关重要。 本项目将开发一种新的体外平台，用于快速分析和抗体中和工程。 我们将建立以高通量直接选择所需抗病毒特性的抗体的方法，包括 中和的广度和效力。我们将为天然配对的抗体重链和重链应用定制平台， 从人类免疫应答中捕获轻链基因，以绘制抗病毒抗体的中和能力 自然感染或疫苗接种引起的，并选择针对相关病毒的广泛抗体保护。 目的一是建立新的抗SARS抗体筛选和工程化的检测技术， CoV-2在最近出现在人类群体中后不断进化。该项目将确定 从恢复期COVID-19患者的免疫反应中获得具有高效力和广度的抗体变体。 我们还将设计有前途的抗体，以增强对各种疾病的中和广度和效力。 SARS-CoV-2毒株。 目的2将建立抗黄病毒抗体的发现和工程策略， 发烧病毒作为一个关键模型系统。黄病毒中的抗体依赖性增强产生有效的抗体 中和是任何基于抗体的临床干预的关键特征。我们会找出有效的中和剂 从接种黄热病病毒疫苗的患者身上提取抗体，并设计改进的中和抗体， 对多种黄热病病毒株的高效力 这项工作将为有效的抗病毒药物发现和抗体工程建立一个新的平台方法。我们 长期目标是开发强大而快速的抗病毒抗体发现平台， 病毒性疾病医疗干预措施的发展进展。

PROJECT SUMMARY A detailed understanding of molecular and cellular adaptive immune responses is critical to accelerate progress in human immunology and drug development. However, available technologies for analyzing antiviral neutralizing antibody responses are slow and impractical for large-scale clinical sample analysis, and can provide limited information on the scope of neutralizing antibody features in human immunity. Importantly, current methods are also unable to engineer antibody molecules to directly improve neutralization potency, which is a major limitation to the discovery of potent and broadly reactive antibody-based interventions for viral diseases. Current methods also cannot engineer broad antibody neutralization against related viruses, which is critical for drug and vaccine development against diverse viral lineages. Important examples include the diverse viral lineages of betacoronaviruses and flaviviruses, where protection against evolved and expanded viral lineages is essential for effective clinical use. This project will develop a new in vitro platform for rapid analysis and engineering of antibody neutralization. We will establish methods to directly select antibodies desired antiviral properties at high throughput, including for neutralization breadth and potency. We will apply a custom platform for natively paired antibody heavy and light chain gene capture from human immune responses to map the neutralization capacity of antiviral antibodies elicited by natural infection or vaccination, and to select for broad antibody protection against related viruses. Aim 1 will establish our new assay techniques for antiviral antibody discovery and engineering against SARS- CoV-2, which is continuously evolving after its recent emergence into human populations. This project will identify antibody variants with high potency and breadth from the immune responses of convalescent COVID-19 patients. We will also engineer promising antibodies for enhanced neutralization breadth and potency against diverse SARS-CoV-2 strains. Aim 2 will establish antiviral antibody discovery and engineering strategies against flaviviruses, using yellow fever virus as a key model system. Antibody-dependent enhancement in flaviviruses makes potent antibody neutralization a critical feature for any antibody-based clinical interventions. We will identify potent neutralizing antibodies from patients vaccinated against yellow fever virus, and engineer improved neutralizing antibodies for high potency against multiple yellow fever virus strains This work will establish a new platform approach for potent antiviral discovery and antibody engineering. Our long-term objectives are to develop robust and rapid antiviral antibody discovery platforms that can accelerate progress in the development of medical interventions for viral diseases.