Neutralizing antibody against SARS-CoV-2 spike in COVID-19 patients, health care workers, and convalescent plasma donors
Cong Zeng, John P. Evans, Rebecca Pearson, Panke Qu, Yi-Min Zheng, Richard T. Robinson, Luanne Hall-Stoodley, Jacob Yount, Sonal Pannu, Rama K. Mallampalli, Linda Saif, Eugene Oltz, Gerard Lozanski, Shan-Lu Liu
Cong Zeng, John P. Evans, Rebecca Pearson, Panke Qu, Yi-Min Zheng, Richard T. Robinson, Luanne Hall-Stoodley, Jacob Yount, Sonal Pannu, Rama K. Mallampalli, Linda Saif, Eugene Oltz, Gerard Lozanski, Shan-Lu Liu
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Resource and Technical Advance COVID-19

Neutralizing antibody against SARS-CoV-2 spike in COVID-19 patients, health care workers, and convalescent plasma donors

Abstract

Rapid and specific antibody testing is crucial for improved understanding, control, and treatment of COVID-19 pathogenesis. Herein, we describe and apply a rapid, sensitive, and accurate virus neutralization assay for SARS-CoV-2 antibodies. The assay is based on an HIV-1 lentiviral vector that contains a secreted intron Gaussia luciferase (Gluc) or secreted nano-luciferase reporter cassette, pseudotyped with the SARS-CoV-2 spike (S) glycoprotein, and is validated with a plaque-reduction assay using an authentic, infectious SARS-CoV-2 strain. The assay was used to evaluate SARS-CoV-2 antibodies in serum from individuals with a broad range of COVID-19 symptoms; patients included those in the intensive care unit (ICU), health care workers (HCWs), and convalescent plasma donors. The highest neutralizing antibody titers were observed among ICU patients, followed by general hospitalized patients, HCWs, and convalescent plasma donors. Our study highlights a wide phenotypic variation in human antibody responses against SARS-CoV-2 and demonstrates the efficacy of a potentially novel lentivirus pseudotype assay for high-throughput serological surveys of neutralizing antibody titers in large cohorts.

Authors

Cong Zeng, John P. Evans, Rebecca Pearson, Panke Qu, Yi-Min Zheng, Richard T. Robinson, Luanne Hall-Stoodley, Jacob Yount, Sonal Pannu, Rama K. Mallampalli, Linda Saif, Eugene Oltz, Gerard Lozanski, Shan-Lu Liu

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Figure 2

Comparison of HIV-1 inGluc pseudotypes bearing C9-tagged spikes of SARS-CoV or WTs.

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Comparison of HIV-1 inGluc pseudotypes bearing C9-tagged spikes of SARS-...
(A) Relative infectivity. Experiments were performed as described as Figure 1, B and C, except that either WT or C9-tagged spikes were used for virus production; n = 6. *P < 0.001, by 2-tailed t test. (B and C) Western blotting analysis of C9-tagged S protein expression in the virus-producing cells (B) and purified viral particles (C). Viral production was carried as described in Figure 1, and viral producer cells were lysed and analyzed by Western blotting using anti-C9, anti-p24, and/or anti–β-actin. (D–F) Virus-producing cells were digested with PBS-5 mM EDTA and incubated with 10 μg/mL sACE2-Fc for 2 hours; cells were washed 3 times, incubated with FITC-labeled anti-human Fc for 45 minutes, and analyzed by flow cytometry. (D) Representative cell populations analyzed for SARS-CoV and SARS-CoV-2; the percentage of positive cells for FITC anti-human Fc was shown. (E) Histogram analysis of virus-producing cells for SARS-CoV and SARS-CoV-2. (F) Relative mean fluorescence intensities of cells expressing indicated spikes; n = 2; no statistical analysis was performed. Note that cells transfected with an empty vector pCIneo served as negative control, the fluorescence intensity of which was set to 1. Data were analyzed as mean ± SD.