Extended Data Fig. 2: hCoV proteins expression is optimized with a GCN4 tag.
From: Converting non-neutralizing SARS-CoV-2 antibodies into broad-spectrum inhibitors
(A) A schematic representation of constructs tested for expression. 6 hCoV spike proteins, either FL or truncated, tested with 4 different C-terminal trimerization domains and tags. ∆C truncations shown in the middle, SARS-CoV-2 spike numbering. (B) linkers tested. For those without Avi tags, the underlined portions were removed. (C) (top) BLI binding responses from isolated Expi supernatants binding to His 1 K octet tips. Higher response corresponds to higher protein expression. (bottom) An anti-his tag dot blot assay of the supernatants tested in the BLI binding. Dot blot shows good correspondence with the BLI binding. Influenza hemagglutinin used as a positive control, mock transfection as a negative control. (D) Yield determined for full length 2 P hCoV constructs containing the GCN4-Avi-His tag from a duplicate experiment of 100 mL transfections, error bars denote standard deviation.
