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

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© 2021 Published by Elsevier Inc.

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Sequential selection of 2B04 and 2H04 escape mutants

(A) Plaque assays were performed to isolate the VSV-SARS-CoV-2-S wild-type, E484A, E484K, and F486S escape mutant on Vero E6 TMPRSS2 cells in the present of the indicated mAb in the overlay. Representative images of three independent experiments are shown.

(B) The surface model of RBD (from PDB 6M0J) is depicted, and contact residues of the SARS-CoV-2 RBD-hACE2 interfaces are colored in brown. 2B04 escape mutants including E484A, E484K, and F486S are indicated in green. Amino acids whose substitution confers resistance to 2H04 in the plaque assays are indicated in lemon.

(C) Wild-type and sequentially identified double mutants were tested for neutralizing activity using a plaque assay with the indicated mAb in the overlay. mAb concentrations added were the same as those used to select the escape mutants. Representative images of two independent experiments are shown.