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 3

Neutralization of SARS-CoV-2 by COVID-19 patient sera.

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Neutralization of SARS-CoV-2 by COVID-19 patient sera. Validation of inG...
Validation of inGluc-based lentiviral pseudotypes using an authentic SARS-CoV-2 US-WA-1 strain. Note that all samples tested here and throughout the studies were blinded before testing. (A) A group of 8 blinded patient sera was tested for neutralization of the SARS-CoV-2 lentiviral S pseudotypes bearing C9-tagged SARS-CoV-2 S or WT. Note that only patient serum sample (sample 8) was tested for the WT spike, the pattern of which almost perfectly overlaps with that of C9-tagged spike; n = 3. The ELISA OD450 values of 8 samples are indicated for each number (cutoff, 0.40). (B) Results of infectious SARS-CoV-2 plaque-reduction neutralization assay for testing of 8 blinded samples. Vero-E6 cells were infected for 3 days with infectious SARS-CoV-2, pretreated with or without the indicated diluted sera. Cells were fixed and stained with 0.25% crystal violet for visualization of plaques; n = 3. Note that the BEI guinea pig antiserum to SARS-CoV did not inhibit SARS-CoV-2 infection. (C) Types of serum or plasma samples did not appear to affect the neutralization pattern generated by inGluc-based lentiviral pseudotypes bearing SARS-CoV-2 spike. Serum, sodium citrate-treated plasma, and EDTA-treated plasma from the same patient were used for SARS-CoV-2 S pseudotype neutralization. Data were analyzed as mean ± SD.