| Application Notes |
The ELISA plates were coated with SARS-CoV-2 antigen or SARS antigen or Middle East respiratory syndrome coronavirus antigen or human coronavirus OC43 antigen at optimal concentrations in carbonate buffer and incubated at 4°C overnight. The next day, unbound antigens were removed by pipetting to avoid the risk of forming aerosols. Non-specific binding was blocked with a solution of PBS with 3% BSA at room temperature for 1h on a shaker. mAb-containing cell culture supernatant or purified mAb preparation was added and incubated at 37°C for 1 h. The non-transfected cell culture supernatant, anti-influenza human monoclonal antibody BS 1A, anti-SARS spike monoclonal antibody CR3022 and convalescent serum were used as antibody controls for each experiment. After incubation, the plate was washed and incubated with HRP-conjugated rabbit anti-human IgG as secondary antibody. After incubation, the plate was washed and developed with 3,3',5,5'-tetramethylbenzidine substrate reagent. Reaction was stopped by 0.5 M hydrochloric acid and the OD was measured at 450 nm on a microplate reader. |
