Abstract
Development of antibody protection during SARS-CoV-2 infection is a pressing question for public health and for vaccine development. We developed highly sensitive SARS-CoV-2-specific antibody and neutralization assays. SARS-CoV-2 Spike protein or Nucleocapsid protein specific IgG antibodies at titers more than 1:100,000 were detectable in all PCR+ subjects (n = 115) and were absent in the negative controls. Other isotype antibodies (IgA, IgG1-4) were also detected. SARS-CoV-2 neutralization was determined in COVID-19 and convalescent plasma at up to 10,000-fold dilution, using Spike protein pseudotyped lentiviruses, which were also blocked by neutralizing antibodies (NAbs). Hospitalized patients had up to 3000-fold higher antibody and neutralization titers compared to outpatients or convalescent plasma donors. Interestingly, some COVID-19 patients also possessed NAbs against SARS-CoV Spike protein pseudovirus. Together these results demonstrate the high specificity and sensitivity of our assays, which may impact understanding the quality or duration of the antibody response during COVID-19 and in determining the effectiveness of potential vaccines.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Adult
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Angiotensin-Converting Enzyme 2 / chemistry
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Angiotensin-Converting Enzyme 2 / immunology
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Angiotensin-Converting Enzyme 2 / metabolism
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Antibodies, Neutralizing / biosynthesis
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Antibodies, Neutralizing / chemistry*
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Antibodies, Viral / biosynthesis
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Antibodies, Viral / chemistry*
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COVID-19 / diagnosis*
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COVID-19 / immunology
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COVID-19 / virology
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Convalescence
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Coronavirus Nucleocapsid Proteins / chemistry*
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Coronavirus Nucleocapsid Proteins / immunology
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Coronavirus Nucleocapsid Proteins / metabolism
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Enzyme-Linked Immunosorbent Assay / methods
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Epitopes / chemistry
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Epitopes / immunology
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Epitopes / metabolism
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Female
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Humans
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Immune Sera / chemistry
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Immunity, Humoral
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Lentivirus / genetics
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Lentivirus / immunology
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Male
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Middle Aged
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Neutralization Tests
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Phosphoproteins / chemistry
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Phosphoproteins / immunology
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Phosphoproteins / metabolism
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Protein Binding
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Receptors, Virus / chemistry
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Receptors, Virus / immunology
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Receptors, Virus / metabolism
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SARS-CoV-2 / drug effects
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SARS-CoV-2 / immunology*
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SARS-CoV-2 / pathogenicity
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Severity of Illness Index
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Spike Glycoprotein, Coronavirus / chemistry*
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Spike Glycoprotein, Coronavirus / immunology
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Spike Glycoprotein, Coronavirus / metabolism
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Survival Analysis
Substances
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Antibodies, Neutralizing
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Antibodies, Viral
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Coronavirus Nucleocapsid Proteins
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Epitopes
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Immune Sera
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Phosphoproteins
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Receptors, Virus
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Spike Glycoprotein, Coronavirus
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nucleocapsid phosphoprotein, SARS-CoV-2
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spike protein, SARS-CoV-2
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ACE2 protein, human
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Angiotensin-Converting Enzyme 2