Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding
Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding
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DOI：
10.1128/msphere.00802-20
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发表时间：
2020-09-01
期刊：
影响因子：
4.8
通讯作者：
Wells, James A.
中科院分区：
文献类型：
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作者：
Byrnes, James R.;Zhou, Xin X.;Wells, James A.
随着严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）继续在世界范围内传播，迫切需要新的检测方法来表征感染的体液反应。在这里，我们提出了一个有效的，竞争性的血清学检测，可以同时确定个人的血清反应性对SARS冠状病毒-2刺突蛋白，并确定比例的抗刺突抗体，阻断相互作用与人血管紧张素转换酶2（ACE 2）所需的病毒进入。在这种方法的基础上使用酶联免疫吸附试验（ELISA），我们提出了天然折叠的病毒刺突蛋白受体结合域（RBD）的抗原通过亲和素-生物素相互作用。然后将血清与可溶性ACE 2-Fc或其更高亲和力的变体竞争，以确定ACE 2阻断性抗RBD抗体的比例。对来自144名SARS-CoV-2患者的血清的评估最终揭示，抗RBD库中的抗体的显著一致性和高比例靶向负责ACE 2接合的表位（83% +/- 11%;在我们最大的组群中50%至107%的信号抑制），进一步强调了定制疫苗以促进此类抗体发展的重要性。重要性随着SARS-CoV的出现和持续传播，2病毒以及相关疾病2019冠状病毒病（COVID-19），迫切需要更好地了解人体如何对病毒产生免疫反应。在这里，我们开发了一种竞争性的SARS-CoV-2血清学检测，可以同时确定一个人是否已经开发出针对SARS-CoV-2刺突蛋白受体结合域（RBD）的抗体，并测量这些抗体的比例，阻断与病毒进入所需的人血管紧张素转换酶2（ACE 2）的相互作用。使用该测定和144份SARSCoV-2患者血清样品，我们发现大多数抗RBD抗体竞争ACE 2结合。这些结果不仅突出了设计疫苗以产生这种阻断抗体的需要，而且还证明了该测定法用于快速筛选患者血清中潜在的中和抗体的实用性。
As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread around the world, there is an urgent need for new assay formats to characterize the humoral response to infection. Here, we present an efficient, competitive serological assay that can simultaneously determine an individual's seroreactivity against the SARS-CoV-2 Spike protein and determine the proportion of anti-Spike antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. In this approach based on the use of enzyme-linked immunosorbent assays (ELISA), we present natively folded viral Spike protein receptor-binding domain (RBD)-containing antigens via avidin-biotin interactions. Sera are then competed with soluble ACE2-Fc, or with a higher-affinity variant thereof, to determine the proportion of ACE2 blocking anti-RBD antibodies. Assessment of sera from 144 SARS-CoV-2 patients ultimately revealed that a remarkably consistent and high proportion of antibodies in the anti-RBD pool targeted the epitope responsible for ACE2 engagement (83% +/- 11%; 50% to 107% signal inhibition in our largest cohort), further underscoring the importance of tailoring vaccines to promote the development of such antibodies.IMPORTANCE With the emergence and continued spread of the SARS-CoV-2 virus, and of the associated disease, coronavirus disease 2019 (COVID-19), there is an urgent need for improved understanding of how the body mounts an immune response to the virus. Here, we developed a competitive SARS-CoV-2 serological assay that can simultaneously determine whether an individual has developed antibodies against the SARS-CoV-2 Spike protein receptor-binding domain (RBD) and measure the proportion of these antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. Using this assay and 144 SARSCoV-2 patient serum samples, we found that a majority of anti-RBD antibodies compete for ACE2 binding. These results not only highlight the need to design vaccines to generate such blocking antibodies but also demonstrate the utility of this assay to rapidly screen patient sera for potentially neutralizing antibodies.