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 mTNFα Target Cells Shows Precision, Accuracy and Linearity.
The ADCC Reporter Bioassay with mTNFα Target Cells measures the activity of the anti-TNFα antibodies infliximab (Panel A) and adalimumab (Panel B). mTNFα 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 infliximab or adalimumab 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 148ng/ml for infliximab and 10.7ng/ml for adalimumab. Fold induction was 4.9 and 2.9 for infliximab and adalimumab, respectively.
A CDC assay measures the activity of the anti-TNFα antibodies infliximab (Panel A) and adalimumab (Panel B). mTNFα Target Cells were added to a 96-well assay plate 18 hours prior to the assay. On the day of assay, 10% CDC-Qualified Human Serum and a titration of infliximab or adalimumab 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. Infliximab gave an EC 50 of 0.73µg/ml, fold induction of 7.7 and specific lysis of 40.2%. With adalimumab, the EC 50 was 0.19µg/ml, the fold induction was 15.7 and the specific lysis was 37.1%.
