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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

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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 5

Evaluations of neutralizing antibody levels in COVID-19 hospitalized inpatients, ICU patients, health care workers, and convalescent plasma donors.

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Evaluations of neutralizing antibody levels in COVID-19 hospitalized inp...
Blinded serum samples were serially diluted and tested for neutralizing activity against lentiviral pseudotypes bearing SARS-CoV-2 S. (A–E) Ranges of neutralizing antibody titer IC50 in the 4 indicated groups (x axis); percent in each study group was plotted (y axis). (F–J) Neutralization curves of 4 different groups, as presented by the relative infectivity of SARS-CoV-2 S pseudotypes in the presence of indicated serum samples. The y axis indicates the relative viral infectivity by setting the viral infectivity without serum to 100%; the x axis indicates dilution fold of serum samples. (K) Correlational analysis of pseudovirus neutralization IC50 and N protein IgG antibody ELISA OD450 values; r = 0.4192, P = 0.0027 as indicated; n = 49. r = 0.4192, P = 0.0027, as indicated by correlation of XY analyses; n = 49. (L) Correlational analysis between pseudovirus neutralization IC50 and age; n = 30. Data were analyzed as mean ± SD.

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