Quantitative Immunological Assays to Assess SARS-CoV-2 Antibodies and Vaccines

在过去的几年中，病毒学实验室与其他内部NIAID实验室和其他外部合作者合作，开发了假病毒中和试验和表达融合前稳定的SARS-CoV-2刺突三聚体（mRNA-1273）的mRNA疫苗。对于中和试验，我们为其他VRC实验室提供了技术和软件帮助，并制定了共享的标准操作规程。由于国际上不断出现新的SARS-CoV-2变异病毒，我们也在不断制作与新出现的变异病毒相对应的各种假病毒。更重要的是，假病毒中和试验也需要进行修改，以适应新的变异病毒，以便试验适当使用。 实验室工作人员正在使用该检测方法评估从COVID-19中恢复的个人以及政府批准的疫苗临床试验中的志愿者的人类血清样本。我们还从COVID-19疾病康复者中分离和鉴定单克隆抗体，评估SARS-CoV-2中和抗体，这是有价值的研究试剂，具有巨大的潜力，可用于治疗和预防不断出现的SARS-CoV-2变体。 通过使用这种强大的检测方法，我们分析了接种疫苗或感染者血液中抗COVID-19抗体的效力和交叉反应性。具体来说，我们已经测量了针对原始或变异SARS-CoV-2病毒的中和抗体，作为mRNA疫苗临床试验的一部分。我们还评估了从COVID-19康复者中分离的单克隆抗体的中和活性，作为我们对病毒免疫的基础研究的一部分，并寻找具有预防或治疗潜力的单克隆抗体。

During the past years, the Virology Laboratory has collaborated with other intramural NIAID labs and other external collaborators to develop a pseudovirus neutralization assay and an mRNA vaccine expressing the prefusion-stabilized SARS-CoV-2 spike trimer (mRNA-1273). For the neutralization assay, we have provided technical and software assistance to other VRC labs, and developed a shared Standard Operating Procedure. Since more new SARS-CoV-2 variant viruses continuously arise in the world, we are continuously making various pseudoviruses corresponding to the emerging variant viruses. And more importantly, the pseudovirus neutralization assay also needed to be modified to adapted to the new variant viruses for the assay appropriate usage. The assay is being used by laboratory staff to evaluate human serum samples from individuals recovering from COVID-19 and from volunteers in the clinical trial of the government approved vaccines. We are also isolating and characterizing monoclonal antibodies from individuals recovering from COVID-19 disease, assessing for SARS-CoV-2 neutralizing antibodies, which are valuable research reagents with great potential for therapeutic and prevention purposes against the continuously emerging SARS-CoV-2 variants. By using this powerful assay, we have analyzed the potency and cross-reactivity of anti-COVID-19 antibodies in the blood of vaccinated or infected individuals. Specifically, we have measured neutralizing antibodies against the original or variant SARS-CoV-2 viruses as part of clinical trials for mRNA vaccines. We have also assessed the neutralization activity of monoclonal antibodies isolated from individuals who have recovered from COVID-19, as part of our basic research on viral immunity, and in search of monoclonal antibodies with preventative or therapeutic potential.