Fig. 6: Generation, biophysical characterization, and functional testing of a humanized mAb clone. | Nature Communications

Fig. 6: Generation, biophysical characterization, and functional testing of a humanized mAb clone.

From: Monospecific and bispecific monoclonal SARS-CoV-2 neutralizing antibodies that maintain potency against B.1.617

Fig. 6

a Schematics of antibody humanization. b SDS-PAGE of purified antibodies of humanized clones (Clone 13A) along with other clones, this experiment had been done for once. c Octet measurement of Clone 13A using the BLI assay. d (Top) Neutralization assay on Clone 2 (n = 3 biological replicates) and a Clone 13A (humanized Clone 2) (n = 3 biological replicates) using WT SARS-CoV-2 Spike pseudotyped HIV-lentivirus carrying an EGFP reporter. (Bottom) Neutralization assay on Clone 2 (n = 2 biological replicates) and a Clone 13A (humanized Clone 2) (n = 3 biological replicates) using B.1.617 variant SARS-CoV-2 Spike pseudotyped HIV-lentivirus carrying an EGFP reporter. e In vitro neutralization of top mAb clones (Clones 2, 6, 13A) along with representative therapeutic antibody clones (RGEN 10933, 10987) against authentic SARS-CoV-2 WA1/2020 and B.1.617.2 (Delta) viruses in BL3 setting (n = 4 biological replicates). f, g Body weight curves of humanized antibody Clone 13 A and placebo (isotype hIgG1) treated hACE2 transgenic mice under lethal challenges of authentic SARS-CoV-2 WA1. f Spider curves of body weight changes of individual mouse plotted separated by group. g Mean ± SEM curves of body weight changes. (Clone 13 A, n = 9, and hIgG1, n = 8 mice, respectively), Clone 13 A vs hIgG1: p = 0.0086. h Survival curves of Clone 13 A antibody and placebo (hIgG1) treated hACE2 transgenic mice under lethal challenges of authentic SARS-CoV-2 (the same experiment in f, g), Clone 13 A vs hIgG1: p < 0.0001, a two-sided statistical test was used to assess statistical significance. i, j Body weight curves of humanized antibody Clone 13 A and placebo (isotype hIgG1) treated hACE2 transgenic mice under lethal challenges of authentic SARS-CoV-2 Delta. i Spider curves of body weight changes of individual mouse plotted separated by group. j Mean ± SEM curves of body weight changes. (Clone 13 A, n = 8, and hIgG1, n = 8 mice, respectively), Clone 13 A vs hIgG1: p < 0.0001. In this figure: Data were shown as mean ± s.e.m. plus individual data points in dot plots. Statistics: Two-way ANOVA was used to assess statistical significance for multi-group curve comparisons; Log-rank test was used to assess statistical significance for survival curve comparisons; unless otherwise noted. The p values are indicated in the plots. Source data and additional statistics for experiments are provided as a Source Data file.

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