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. 2022 Jan 5;12:790415. doi: 10.3389/fimmu.2021.790415

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Copyright © 2022 Vanderheijden, Stevaert, Xie, Ren, Barbezange, Noppen, Desombere, Verhasselt, Geldhof, Vereecke, Stroobants, Oh, Vanhee, Naesens and Nauwynck

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Virus-neutralizing properties of peptide-purified human antibodies and parent serum 20Hu384. (A) The virus-neutralizing activity was determined by seroneutralization (SN) assay against FeCV in feline enterocytes, HCoV-OC43 in HRT-18 cells, and SARS-CoV-2 in VeroE6 cells. The limit of detection (LOD) was a 1:8 (FeCV) or 1:40 dilution (HCoV-OC43 and SARS-CoV-2). (B, C) Evaluation of the pAbs (starting dilution: 1:2) in a SARS-CoV-2 neutralization assay in Calu-3 cells. (B) Evaluation based on viral load quantification at 24 h p.i. The three right panels show the negative control (elution buffer); parent human serum #20Hu384; and positive control antibody (S1-mAb: SARS-CoV-2 neutralizing anti-S1 antibody). Data are the mean ± SEM of two independent experiments, performed in duplicate. The Y-axis shows the fold reduction in viral RNA versus untreated virus control, based on qRT-PCR analysis of the supernatants at 24 h p.i. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001, ****P ≤ 0.0001 (multiple unpaired t-test with Holm-Šídák correction; treated sample versus untreated virus control). (C) In parallel, virus replication was quantified by immunofluorescence staining for dsRNA at 72 h p.i. The graphs represent the percentage neutralization in function of antibody dilution or concentration.