Promega, JA6705, CD28 Bioassay

Assay Performance Data
Accuracy and precision are critical to bioassays and the development of biologics. The CD28 Bioassay is both accurate and precise. Accuracy is a measure of how close results are to a target or reference value. Precision is a measure of repeatability. 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 CD28 Bioassay Shows Precision, Accuracy and Linearity.
The CD28 Bioassay reflects the mechanism of action (MOA) and show specificity of biologics designed to activate CD28.
CD28 Effector Cells were incubated with FcγRIIb aAPC/CHO-K1 Cells in the presence of serial titrations of agonist antibodies as indicated. After a 5-hour induction, Bio-Glo™ Reagent was added and luminescence quantified using the GloMax® Discover System. Data were generated using thaw-and-use cells.
FcγRIIb aAPC/CHO-K1 Cells were added to a 96-well assay plate 18 hours prior to the assay. On the day of the assay, a titration of Control Ab, Anti-CD28 (Cat.# K1231), and CD28 Effector Cells was added. After a 5-hour induction at 37°C, Bio-Glo™ Reagent was added and luminescence measured using the GloMax® Discover System.