The iLite® ADCC Target CD20 (+) Assay Ready Cells can be used together with matched iLite® ADCC Effector (V) and iLite® ADCC Target CD20 (-) Assay Ready Cells for the quantification ADCC activity.
Application notes for the following assays are available:
• 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 has existed since the discovery of the ADCC mechanism. Rituximab, one of the first of such drugs approved, is a chimeric monoclonal antibody targeting CD20, a surface antigen primarily found on B-cells. The drug was approved by the FDA in 1997 for treatment of chemotherapy resistant Non-Hodgkin B-cell lymphomas, and has since also been approved in Europe for different inflammatory indications (2-4).
In addition, other anti-CD20 monoclonal antibodies have been developed, such as ocrelizumab, a humanized (90-95%) antibody, and ofatumumab, a fully human monoclonal. In addition, efforts are being made to enhance the ADCC activity of the antibodies by engineering the glycan patterns of the constant region, such as in obinutuzumab (5).
>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).