Description
The Assay Principle
Representation of co-stimulatory bioassays (CD40, OX40, GITR or 4-1BB) with FcγRIIb-independent and FcγRIIb-dependent antibodies. Panel A. Assay with FcγRIIb-independent antibody. The bioassay consists of one engineered cell line: CD40, OX40, GITR or 4-1BB Effector Cells. In the absence of agonist antibody or ligand, the CD40, OX40, GITR or 4-1BB receptor is not activated and luminescence signal is low. The addition of agonist antibody or ligand induces the CD40, OX40, GITR or 4-1BB pathway-activated luminescence, which can be detected in a dose-dependent manner by addition of Bio-Glo™ Reagent and quantitation with a luminometer. Panel B. Assay with FcγRIIb-dependent antibody. The bioassay consists of two engineered cell lines: CD40, OX40, GITR or 4-1BB Effector Cells and FcγRIIb CHO-K1 Cells. In the presence of FcγRIIb CHO-K1 Cells, the anti-CD40, -OX40, -GITR or 4-1BB antibody is cross-linked, thereby inducing CD40, OX40, GITR or 4-1BB pathway-activated luminescence, which can be detected in a dose-dependent manner by addition of Bio-Glo™ Reagent and quantitation with a luminometer.
