Development and characterization of engineered therapeutic antibodies against SARS-CoV-2

项目摘要/摘要 这份R01题为《针对SARS冠状病毒的工程化治疗性抗体的研制与鉴定-- 2“，基于我们在表征病毒-宿主因子相互作用方面的项目基础设施、专业知识和经验 在负链RNA病毒中。自从起源于中国以来，SARS-CoV-2迅速传播，现在是一种 全球大流行。值得关注的是，人类在免疫方面是幼稚的，而且没有可用的 治疗。在美国，这种疾病已经使一些州的医疗系统不堪重负，并有 严重的连锁反应，加剧了对其他疾病的护理标准。在撰写本文时，已有近5家公司 已有100万例和160,000例死亡归咎于新冠肺炎。病毒在肺部复制， 导致一种严重的呼吸道疾病新冠肺炎，这种疾病在2%的病例中是致命的。中和抗体(NAB) 由自然感染或疫苗产生的已知可以控制许多感染和目前的早期研究 新冠肺炎大流行，包括测试恢复期血浆疗法的研究，是有希望的。这些研究 强调基于NAB的治疗的潜在意义。而长期以来，免疫球蛋白抗体的格式一直是最多的 广泛使用的形式，早期的研究，包括我们自己的研究表明，额外的多价形式的NaB可能 要更有效率。这为开发nabs提供了一种创新的方法，同时从以下方面获得潜在好处 在较低剂量下有效中和，并降低出现抗药性突变的可能性。SARS冠状病毒- 2是一种单链、无节段、有包膜的RNA病毒。病毒感染需要尖峰的相互作用 糖蛋白受体结合域(RBD)与宿主受体ACE2结合。在这里，我们将在新开发的基础上 和已建立的方法，通过我们的工作优化了其他系统以产生抗体 利用噬菌体展示技术靶向SPEK和SPEK RBD，并表征其物理特性。我们 将设计出效价更高的抗体，并在体外中和试验和体内试验中测试效力 在小鼠模型中的疗效。在完成后，我们希望提供创新和独特的多价NAB引线 具有独特的特性，可与同类最好的免疫球蛋白药物相媲美。

Project Summary/Abstract This R01 entitled, “Development and characterization of engineered therapeutic antibodies against SARS-CoV- 2”, builds on our project infrastructure, expertise, and experience in characterizing viral-host factor interactions in negative strand RNA viruses. Since originating in China, SARS-CoV-2 has since rapidly spread and is now a global pandemic. Significant concerns are that humans are immunologically naïve, and there are no available therapies. In the US, the disease has already overwhelmed the healthcare system in some states and have a serious knock-on effect in exacerbating the standard of care for other diseases. At the time of writing, nearly 5 million cases and >160,000 deaths have been attributed to COVID-19. The virus replicates in the lungs and causes a severe respiratory disease, COVID-19, which is fatal in >2% of cases. Neutralizing antibodies (nAbs) generated by natural infection or vaccines is known to control many infections and early studies in the current COVID-19 pandemic, including studies to test convalescent plasma treatments, are promising. These studies highlight the potential significance of nAb-based therapy. While IgG format of nAbs have long been the most extensively used format, early studies, including our own suggest that additional multivalent formats of nAbs may be more effective. This provides an innovative method to develop nAbs while acquiring potential benefits from effective neutralization at lower doses and lower likelihood of the emergence of resistance mutants. SARS-CoV- 2 is a single stranded, non-segmented, enveloped RNA virus. Viral infection requires interaction of the spike glycoprotein receptor binding domain (RBD) to the host receptor ACE2. Here, we will build on newly developed and established approaches that have been optimized through our work other systems to generate antibodies targeting spike and spike RBD using phage display technology and characterize their physical properties. We will engineer antibodies with increased valency and test for potency in in vitro neutralization assays and in vivo efficacy in a mouse model. At the completion, we expect to provide innovative and unique multivalent nAb leads with unique characteristics that will rival the best in class IgG drugs.