ADCC Reporter Bioassay for Antibodies

The ADCC reporter bioassay core kit was purchased from Promega and used according to manufacturer’s instructions and as reported (38 (link)). Briefly, confluent monolayers of MDCK cells in a white tissue culture treated 96-well plate and infected with Bel/09 at an MOI of 1. Twenty-four hours later, infected cells were used as target cells in the ADCC reporter assay. Human monoclonal antibodies or the human IgG1 isotype control (BioXCell) were added in triplicate in a 3-fold dilution series starting at 100 μg/ml. ADCC bioassay effector cells (Jurkat V variant cells) were added, and plates were incubated at 37°C for 6 h. Bio-Glo luciferase assay reagent was added, and luminescence was measured. Fold induction was calculated from controls with buffer alone, and the 50% effective concentration (EC₅₀) was determined in GraphPad Prism.