Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding.,mSphere

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Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding.
mSphere ( IF 3.7 ) Pub Date : 2020-09-16 , DOI: 10.1128/msphere.00802-20
James R Byrnes ₁ , Xin X Zhou ₁ , Irene Lui ₁ , Susanna K Elledge ₁ , Jeff E Glasgow ₁ , Shion A Lim ₁ , Rita P Loudermilk _{2,

3} , Charles Y Chiu _{4,

5} , Taia T Wang _{6,

7,

8} , Michael R Wilson _{2,

3} , Kevin K Leung ₁ , James A Wells _{6,

9,

10}

Affiliation

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.
Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, California, USA.
Department of Neurology, University of California, San Francisco, San Francisco, California, USA.
Department of Laboratory Medicine, University of California, San Francisco, San Francisco, California, USA.
Department of Medicine, University of California, San Francisco, San Francisco, California, USA.
Chan Zuckerberg Biohub, San Francisco, California, USA.
Department of Medicine, Stanford University Medical School, Stanford, California, USA.
Department of Microbiology and Immunology, Stanford University Medical School, Stanford, California, USA.
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.

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.

中文翻译：

竞争性 SARS-CoV-2 血清学显示，大多数针对尖峰受体结合域的抗体都在竞争 ACE2 结合。

随着严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 继续在世界范围内传播，迫切需要新的检测形式来表征对感染的体液反应。在这里，我们提出了一种有效的、竞争性的血清学检测方法，可以同时确定个体对 SARS-CoV-2 Spike 蛋白的血清反应性，并确定阻断与所需的人血管紧张素转换酶 2 (ACE2) 相互作用的抗 Spike 抗体的比例用于病毒进入。在这种基于使用酶联免疫吸附测定 (ELISA) 的方法中，我们通过亲和素-生物素相互作用呈现天然折叠的病毒穗蛋白受体结合域 (RBD) 包含抗原。然后血清与可溶性 ACE2-Fc 或其更高亲和力的变体竞争，确定ACE2阻断抗RBD抗体的比例。对 144 名 SARS-CoV-2 患者的血清的评估最终显示，抗 RBD 池中非常一致且高比例的抗体靶向负责 ACE2 参与的表位（83% ± 11%；50% 至 107% 的信号抑制）我们最大的队列），进一步强调了定制疫苗以促进此类抗体开发的重要性。

更新日期：2020-09-16

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