Normalization of cell counts, serum matrix effects or lysis of the effector cells by the target cells is obtained by a second reporter gene, a NanoLuc Luciferase reporter gene construct, under control of a constitutive promotor.
The iLite® ADCC Effector (V) Assay Ready Cells can be used together with matched iLite ADCC Target CD20 (+) and iLite® ADCC Target CD20 (-) Assay Ready Cells for the quantification ADCC activity. Please see:
• Quantification of anti-CD20 ADCC activity (E-229-GB)
Antibody-dependent cell-mediated cytotoxicity (ADCC) is a mechanism whereby pathogenic cells are lysed by lymphocytes, most often Natural Killer (NK) cells. The mechanism involves binding of antibodies to surface antigens on the pathogen. Crosslinking of these antibodies to NK cells through the binding of the Fc-portion to Fc receptors on the NK cells leads to activation of the NK cell and formation of an immune synapse with the pathogenic cell. The NK cell releases cytotoxic granules containing granzymes and perforin into the synapse, leading to apoptosis of the targeted cell (1).
The idea of employing ADCC to destroy dysfunctional cells by treating patients with antibodies which induce this mechanism has existed since the discovery of the ADCC mechanism. The first monoclonal antibody for treating cancer to be FDA approved was Rituximab which in part utilizes the ADCC mechanism to destroy cancer cells expressing CD20. Induction of ADCC through monoclonal antibodies is also utilized in treating autoimmune diseases (2, 3).
>250 µL of iLite® Assay Ready Cells suspended in RPMI 1640 with 20% heat inactivated fetal bovine serum (FBS), mixed 1:1 with cryoprotective medium from Lonza (Cat. No 12-132A).