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

被引:53
|
作者
Byrnes, James R. [1 ]
Zhou, Xin X. [1 ]
Lui, Irene [1 ]
Elledge, Susanna K. [1 ]
Glasgow, Jeff E. [1 ]
Lim, Shion A. [1 ]
Loudermilk, Rita P. [2 ,3 ]
Chiu, Charles Y. [4 ,5 ]
Wang, Taia T. [6 ,7 ,8 ]
Wilson, Michael R. [2 ,3 ]
Leung, Kevin K. [1 ]
Wells, James A. [1 ,6 ,9 ]
机构
[1] Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA
[2] Univ Calif San Francisco, Weill Inst Neurosci, San Francisco, CA 94143 USA
[3] Univ Calif San Francisco, Dept Neurol, San Francisco, CA 94143 USA
[4] Univ Calif San Francisco, Dept Lab Med, San Francisco, CA 94143 USA
[5] Univ Calif San Francisco, Dept Med, San Francisco, CA 94143 USA
[6] Chan Zuckerberg Biohub, San Francisco, CA 94158 USA
[7] Stanford Univ, Dept Med, Med Sch, Stanford, CA 94305 USA
[8] Stanford Univ, Dept Microbiol & Immunol, Med Sch, Stanford, CA 94305 USA
[9] Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94143 USA
基金
美国国家卫生研究院; 美国国家科学基金会;
关键词
COVID-19; SARS-CoV-2; angiotensin-converting enzyme 2; immunoserology; neutralizing antibodies; receptor-binding domain; serology;
D O I
10.1128/mSphere.00802-20
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页数:11
相关论文
共 50 条
  • [11] Small Molecules Targeting SARS-CoV-2 Spike Glycoprotein Receptor-Binding Domain
    Rodriguez, Yoel
    Cardoze, Scarlet Martinez
    Obineche, Onyinyechi W.
    Melo, Claudia
    Persaud, Ashanna
    Romero, Jose A. Fernandez
    ACS OMEGA, 2022, 7 (33): : 28779 - 28789
  • [12] Discovery of Aptamers Targeting the Receptor-Binding Domain of the SARS-CoV-2 Spike Glycoprotein
    Song, Yanling
    Song, Jia
    Wei, Xinyu
    Huang, Mengjiao
    Sun, Miao
    Zhu, Lin
    Lin, Bingqian
    Shen, Haicong
    Zhu, Zhi
    Yang, Chaoyong
    ANALYTICAL CHEMISTRY, 2020, 92 (14) : 9895 - 9900
  • [13] Binding of SARS-CoV-2/SARS-CoV spike protein with human ACE2 receptor
    Koirala, Rajendra P.
    Thapa, Bidhya
    Khanal, Shyam P.
    Powrel, Jhulan
    Adhikari, Rajendra P.
    Adhikari, Narayan P.
    JOURNAL OF PHYSICS COMMUNICATIONS, 2021, 5 (03):
  • [14] Endocytosis of the receptor-binding domain of SARS-CoV spike protein together with virus receptor ACE2
    Wang, Shunxin
    Guo, Feng
    Liu, Kangtai
    Wang, Hongliang
    Rao, Shuan
    Yang, Peng
    Jiang, Chengyu
    VIRUS RESEARCH, 2008, 136 (1-2) : 8 - 15
  • [15] Synthetic Homogeneous Glycoforms of the SARS-CoV-2 Spike Receptor-Binding Domain Reveals Different Binding Profiles of Monoclonal Antibodies
    Ye, Farong
    Zhao, Jie
    Xu, Peng
    Liu, Xinliang
    Yu, Jing
    Shangguan, Wei
    Liu, Jiazhi
    Luo, Xiaosheng
    Li, Cheng
    Ying, Tianlei
    Wang, Jing
    Yu, Biao
    Wang, Ping
    ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2021, 60 (23) : 12904 - 12910
  • [16] Biophysics of SARS-CoV-2 spike protein's receptor-binding domain interaction with ACE2 and neutralizing antibodies: from computation to functional insights
    da Silva, Fernando Luis Barroso
    Paco, Karen
    Laaksonen, Aatto
    Ray, Animesh
    BIOPHYSICAL REVIEWS, 2025,
  • [17] In silico design of ACE2 mutants for competitive binding of SARS-CoV-2 receptor binding domain with hACE2
    Lim, Theam Soon
    Choong, Yee Siew
    PHYSICAL SCIENCES REVIEWS, 2022, : 3595 - 3603
  • [18] Deep Mutational Scanning of SARS-CoV-2 Receptor Binding Domain Reveals Constraints on Folding and ACE2 Binding
    Starr, Tyler N.
    Greaney, Allison J.
    Hilton, Sarah K.
    Ellis, Daniel
    Crawford, Katharine H. D.
    Dingens, Adam S.
    Navarro, Mary Jane
    Bowen, John E.
    Tortorici, M. Alejandra
    Walls, Alexandra C.
    King, Neil P.
    Veesler, David
    Bloom, Jesse D.
    CELL, 2020, 182 (05) : 1295 - +
  • [19] Effect of An 84-bp Deletion of the Receptor-Binding Domain on the ACE2 Binding Affinity of the SARS-CoV-2 Spike Protein: An In Silico Analysis
    Kemenesi, Gabor
    Toth, Gabor Endre
    Bajusz, David
    Keseru, Gyorgy M.
    Terhes, Gabriella
    Burian, Katalin
    Zeghbib, Safia
    Somogyi, Balazs A.
    Jakab, Ferenc
    GENES, 2021, 12 (02) : 1 - 10
  • [20] Atomistic insights into the binding of SARS-CoV-2 spike receptor binding domain with the human ACE2 receptor: The importance of residue 493
    Carter, Camryn
    Airas, Justin
    Parish, Carol A.
    JOURNAL OF MOLECULAR GRAPHICS & MODELLING, 2023, 118