PA22-176, SBIR, Phase I, Development of a SARS-CoV-2 emerging variant infectivity and immune evasion panel to quantify the efficacy of vaccine booster induced neutralizing antibodies.

摘要 随着COVID19大流行和疫苗部署的继续，迫切需要监测新出现的情况 其传染性和逃脱先前感染和疫苗的能力的变体可诱导中和抗体。 目前，还没有办法快速量化新出现的变异体的传染性和免疫逃避能力。这个 量化病毒感染性的标准包括使用收获的病毒分离株并进行定量 斑块分析，繁琐而耗时。此外，传染性SARS-CoV-2的使用需要 BSL-3设施的高水平遏制限制了其在普通临床和研究实验室中的应用。 伪病毒已被广泛用于模拟SARS-CoV-2的感染性和抗体中和。 伪病毒，如来自慢病毒和水泡性口炎病毒的病毒，可以模拟进入过程。 SARS-CoV-2。然而，这些假病毒粒子主要由非冠状病毒结构蛋白组成，需要 2-3天才能产生结果。最近，一种新的杂交甲病毒-SARS-CoV-2伪病毒粒子(HA-CoV-2)已经 由P.I.(Hetrick)开发，用于快速(3-6小时)和准确地量化病毒感染性和 对中和抗体的敏感性。 HA-CoV-2颗粒是一种不可复制的SARS-CoV-2病毒样颗粒 (VLP)仅由SARS-CoV-2结构蛋白(S、M、N和E)和来自RNAs的报告基因组组成 来自一种快速表达的甲型病毒载体。我们的初步研究表明，HA-CoV-2检测 可以快速量化接种过疫苗和之前感染过的人中和抗体的差异 针对特定的SARS-CoV-2变种。Virongy最近从George Mason那里获得了HA-CoV-2技术的许可 并计划开发带有代表SARS的HA-CoV-2颗粒板的商业试剂盒。 目前流通中的CoV-2变种具有最大的传染性和免疫逃逸潜力。像病毒一样 变种不断涌现，迫切需要监测传染性和评估疫苗的有效性。 针对新兴变种的疫苗。HA-CoV-2系统将为快速和准确地 病毒变异传染性和中和抗体反应的量化。我们建议发展一个房委会- 以CoV-2假病毒为基础的检测感染性和中和抗体反应的小组有两个特定目的。 具体目标1是筛选循环中最流行的50种SARS-CoV-2变异刺突蛋白并鉴定 使用HA-CoV-2伪病毒平台的传染性最高的新兴变种。具体目标2是 对循环中的每个冠状病毒变异株进行抗体中和试验，并测定其敏感性。 疫苗和感染的每一种变异体都能诱导抗血清和恢复期血浆。在这项研究中，世卫组织 抗SARS-CoV-2抗体的国际标准和参照小组将与一个 标准重组RBD抗体。在初步筛选之后，将选择前5个变种进行开发 为疫苗和助推器开发人员提供的商业小组试剂盒，用于评估他们的预防药物的效果。作为一部分 在这一筛选过程中，Virongy计划与约翰·霍普金斯大学和其他非营利性公共机构合作 卫生组织提供这项研究中获得的所有数据。HA-CoV-2伪病毒目前 在Virongy的公司网站(Environmental gy.com)上销售，仅用于研究用途。

Summary As the COVID19 pandemic and vaccine deployment continue there is an urgent need to monitor emerging variants for their infectivity and ability to escape both prior infection and vaccine induced neutralizing antibodies. Currently, there is no way to rapidly quantify emerging variants infectivity and immune evading capacity. The standard for quantifying viral infectivity involves the use of harvested viral isolates and performing quantitative plaque assays, which are tedious and time consuming. Additionally, the use of infectious SARS-CoV-2 requires high-level containment in BSL-3 facilities that limits its application in common clinical and research laboratories. Pseudoviruses have been widely used to model SARS-CoV-2 infectivity and antibody neutralization. Pseudoviruses, such as those derived from lentivirus and vesicular stomatitis virus, can mimic the entry process of SARS-CoV-2. However, these pseudovirions consist of mostly non-coronavirus structural proteins and require 2-3 days to generate results. Recently, a novel hybrid alphavirus-SARS-CoV-2 pseudovirion (HA-CoV-2) has been developed by the P.I. (Hetrick) for rapid (3-6 hours) and accurate quantification of viral infectivity and sensitivity to neutralizing antibodies. The HA-CoV-2 particle is a non-replicating SARS-CoV-2 virus-like particle (VLP) composed of only SARS-CoV-2 structural proteins (S, M, N, and E) and a RNA reporter genome derived from a fast expressing alphavirus vector. Our preliminary studies have demonstrated that the HA-CoV-2 assay can rapidly quantify the differences in neutralizing antibodies from vaccinated and previously infected individuals to specific SARS-CoV-2 variants. Virongy has recently licensed the HA-CoV-2 technology from George Mason University, and plans to develop commercial kits with panels of HA-CoV-2 particles that represent the SARS- CoV-2 variants currently in circulation with the greatest infectivity and potential for immune escape. As viral variants continue to emerge there is an urgent need to monitor the infectivity and evaluate the effectiveness of the vaccines on emerging variants. The HA-CoV-2 system would provide a robust platform for rapid and accurate quantification of viral variant infectivity and the neutralizing antibody response. We propose to develop a HA- CoV-2 pseudovirus-based panel to examine infectivity and neutralizing antibody response with two specific aims. Specific Aim 1 is to screen the 50 most prevalent SARS-CoV-2 variant spike proteins in circulation and identify the emerging variants with the highest infectivity using the HA-CoV-2 pseudovirus platform. Specific aim 2 is to conduct antibody neutralization assays of each COVID variant in circulation and determine the sensitivity of each variant to both vaccine and infection induced anti-serum and convalescent plasma. For this study the WHO International Standard and Reference Panel for anti-SARS-CoV-2 antibody will be screened along with a standard recombinant RBD antibody. Following the initial screening the top 5 variants will be selected to develop a commercial panel kit for vaccine and booster developers to evaluate the efficacy of their prophylactics. As part of this screening process Virongy plans to collaborate with John Hopkins university and other non-profit public health organizations to provide all the data acquired in this study. The HA-CoV-2 pseudovirus are currently marketed on Virongy’s company website (virongy.com) for research use only.