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 mVEGF Target Cells Shows Precision, Accuracy and Linearity.
The ADCC Bioassay Effector Cells with mVEGF Target Cells measures the activity of the anti-VEGF antibody bevacizumab. mVEGF 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, Propagation Model (Cat.# G7102), ADCC assay buffer and a titration of bevacizumab were added. After a 24-hour induction at 37°C, Bio-Glo™ 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 0.72µg/ml and fold induction was 3.8.
A CDC assay measures the activity of the anti-VEGF antibody bevacizumab. mVEGF 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 bevacizumab in CDC assay buffer were added. After a 6-hour induction at 37°C, CytoTox-Glo™ 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 for bevacizumab was 2.94µg/ml, fold induction was 6.6 and specific lysis was 25.6%.
