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10.1128/mSphere.00802-20

http://scihub22266oqcxt.onion/10.1128/mSphere.00802-20
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32938700!7494835!32938700
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suck abstract from ncbi


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pmid32938700      mSphere 2020 ; 5 (5): ä
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  • Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding #MMPMID32938700
  • Byrnes JR; Zhou XX; Lui I; Elledge SK; Glasgow JE; Lim SA; Loudermilk RP; Chiu CY; Wang TT; Wilson MR; Leung KK; Wells JA
  • mSphere 2020[Sep]; 5 (5): ä PMID32938700show ga
  • 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 SARS-CoV-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.
  • |Angiotensin-Converting Enzyme 2[MESH]
  • |Antibodies, Neutralizing/*immunology[MESH]
  • |Antibodies, Viral/*immunology[MESH]
  • |Antigens, Viral/immunology[MESH]
  • |Betacoronavirus/*immunology[MESH]
  • |Binding Sites/immunology[MESH]
  • |COVID-19[MESH]
  • |Coronavirus Infections/prevention & control[MESH]
  • |High-Throughput Screening Assays/methods[MESH]
  • |Humans[MESH]
  • |Pandemics/prevention & control[MESH]
  • |Peptidyl-Dipeptidase A/*immunology[MESH]
  • |Pneumonia, Viral/prevention & control[MESH]
  • |Protein Binding[MESH]
  • |Protein Domains/immunology[MESH]
  • |SARS-CoV-2[MESH]
  • |Serologic Tests/*methods[MESH]


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