Rbd of sars cov 2 spike protein

For binding and quantification of serum and BAL antibodies, NUNC MaxiSorp 96-well plates (Thermo Scientific) were coated with 1 μg/mL RBD of SARS-CoV-2 spike protein (Genscript) in PBS overnight at 4°C. The following day, plates were washed with PBS-0.5% Tween 20 and blocked with PBS-10% fetal bovine serum. Next, the plates were again washed and subsequently incubated with diluted mouse sera for 2 h. Following primary serum incubation, plates were incubated with goat anti-mouse IgG-HRP (Bethyl) for 1 h at room temperature. TMB (Thermo) was used to develop the binding signal. Upon stopping with 1N sulfuric acid, plates were read with the BioTek Synergy 2 plate reader (BioTek) at 450nm. Data were subsequently exported to Microsoft Excel and analyzed in graphpad prism version 8.

For binding and quantification of serum and BAL antibodies, NUNC MaxiSorp 96-well plates (Thermo Scientific) were coated with 1μg/mL RBD of SARS-CoV-2 spike protein (Genscript) in PBS overnight at 4°C. The following day, plates were washed with PBS-0.5% Tween 20 and blocked with PBS-10% fetal bovine serum. Next, the plates were again washed and subsequently incubated with diluted mouse sera for two hours. Following primary serum incubation, plates were incubated with goat anti-mouse IgG-HRP (Bethyl) for one hour at room temperature. TMB (Thermo) was used to develop the binding signal. Upon stopping with 1N sulfuric acid, plates were read with the BioTek Synergy 2 plate reader (BioTek) at 450nm. Data were subsequently exported to Microsoft Excel and analyzed in graphpad prism version 8.