Fig. 2: Cross-neutralization capacity of mAbs 28D9 and 1.6C7 and mechanism of action. | Nature Communications

Fig. 2: Cross-neutralization capacity of mAbs 28D9 and 1.6C7 and mechanism of action.

From: A conserved immunogenic and vulnerable site on the coronavirus spike protein delineated by cross-reactive monoclonal antibodies

Fig. 2: Cross-neutralization capacity of mAbs 28D9 and 1.6C7 and mechanism of action.The alternative text for this image may have been generated using AI.

a Antibody-mediated neutralization of infection of luciferase-encoding VSV particles pseudotyped with spike proteins of MERS-CoV, SARS-CoV, SARS-CoV-2 and HCoV-OC43. Pseudotyped VSV particles pre-incubated with antibodies at indicated concentrations were used to infect VeroCCL81 cells (MERS-S pseudotyped VSV), VeroE6 cells (SARS-S and SARS2-S pseudotyped VSV) or HRT-18 cells (OC43-S pseudotyped VSV) and luciferase activities in cell lysates were determined at 20 h post transduction to calculate infection (%) relative to non-antibody-treated controls. The average ± SD (n ≥ 6) from at least two independent experiments performed is shown. Iso-CTRL: an anti-Strep-tag human monoclonal antibody was used as an antibody isotype control. The IC50 and IC90 values were shown. Source data are provided as a Source Data file. b Antibody-mediated neutralization of MERS-CoV, SARS-CoV and SARS-CoV-2 infection. Neutralization of authentic viruses was performed using a plaque reduction neutralization test (PRNT) on VeroCCL81 cells (MERS-CoV) or VeroE6 (SARS-CoV and SARS-CoV-2) as described earlier74,75. The experiment was performed with triplicate samples, the average ± SD is shown. The IC50 and IC90 values were shown. Source data are provided as a Source Data file. c ELISA-based receptor-binding inhibition assay. MERS-Secto pre-incubated with serially diluted mAbs was added to ELISA plates coated with soluble human DPP4. The binding of MERS-Secto to DPP4 was detected using an HRP-conjugated antibody recognizing the C-terminal Strep-tag on MERS-Secto. Data points represent the average ± SD, for n = 3 replicates from two independent experiments. Source data are provided as a Source Data file. d Cell-cell fusion inhibition assay. Huh-7 cells—transfected with plasmid expressing (GFP-tagged) MERS-CoV S were pre-incubated in the presence or absence of 1.6C7 and 28D9, or an irrelevant iso-type control antibody (Iso-CTRL), and then treated with trypsin to activate the membrane fusion function of the MERS-CoV S protein. The formation of MERS-S mediated syncytia was visualized by fluorescence microscopy. Merged images of MERS-S-GFP expressing cells (green) and DAPI-stained cell nuclei (blue) are shown. The experiment was performed twice, data from a representative experiment is shown. Source data are provided as a Source Data file.

Back to article page