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. 2021 Jan 27;29(3):477–488.e4. doi: 10.1016/j.chom.2021.01.014

Figure 1.

Figure 1

VSV-SARS-CoV-2 escape mutant isolation

(A) Outline of escape mutant selection experiment. 2B04 and a control anti-influenza virus mAb were tested for neutralizing activity against VSV-SARS-CoV-2. The concentration of 2B04 added in the overlay completely inhibited viral infection (middle panel). Data are representative of two independent experiments. Plaque assays were performed to isolate the VSV-SARS-CoV-2 escape mutant on Vero E6 TMPRSS2 cells (red arrow indicated). Plaque assays with 2B04 in the overlay (bottom plaque in the right panel); plaque assays without Ab in the overlay (top plaque in the right panel). Data are representative of three independent experiments.

(B) Schematic of S gene, which underwent Sanger sequencing to identify mutations (left panel). For validation, each VSV-SARS-CoV-2 mutant was tested in plaque assays with or without 2B04 in the overlay on Vero cells (right panel). Representative images of two independent experiments are shown.

(C) Neutralizing mAbs. a The order of immunogens used to immunize the mice, as described in STAR methods. b Neutralization of SARS-CoV-2 by each mAb was assessed by focus-reduction neutralization test. The half-maximal effective concentration (EC50 value) was determined by nonlinear regression. Results are the geometric mean from three to four independent experiments. c mAb was identified as mouse IgG1 and expressed as human IgG1.