Description
Assay Performance Data
Accuracy and precision are critical to bioassays and the development of biologics. This assay was developed and prequalified according to ICH guidelines for accuracy and precision, giving it the performance characteristics required for many applications of a bioassay used for antibody drug discovery research and development. Accuracy: Measure of how close results are to a reference or "target" value. IntraLab Precision: Measure of repeatability from multiple assays of the same homogeneous sample under the same operating conditions over a short time (same analyst; same lab; same day). Intermediate Precision: Measure of repeatability of multiple assays of the same homogeneous sample with different analysts or equipment or on different days. Linearity: Ability of the assay to provide measured values that are proportional to the concentration of the sample.
The ADCC Reporter Bioassay using mRANKL Target Cells Shows Precision, Accuracy and Linearity.
The ADCC Reporter Bioassay with mRANKL Target Cells measures the activity of the anti-RANKL antibody denosumab-IgG1. mRANKL Target Cells were added to a 96-well assay plate 18 hours prior to the assay. On the day of the assay, ADCC Effector Cells and a titration of denosumab-IgG1 were added. After a 24-hour induction at 37°C, Bio-Glo™ Luciferase Reagent was added and luminescence measured using the GloMax® Discover System. Data were fitted to a four-parameter logistic curve using GraphPad Prism® software. The EC 50 was 69ng/ml for denosumab-IgG1 and fold induction was 10.2.
A CDC assay measures the activity of the anti-RANKL antibody denosumab-IgG1. mRANKL Target Cells were added to a 96-well assay plate 18 hours prior to the assay. On the day of assay, 10% Normal Human Serum Complement and a titration of denosumab-IgG1 were added. After a 6-hour induction at 37°C, CytoTox-Glo™ Cytotoxicity Reagent was added and luminescence measured using the GloMax® Discover System. Data were fitted to a four-parameter logistic curve using GraphPad Prism® software. For denosumab-IgG1, the EC 50 was 261ng/ml, fold induction was 11.0 and specific lysis was 53%.
