iLite® Effector (V) Assay Ready Cells :
- Storage: -80°C, Cells should be used within 30 min of thawing.
- Contents: >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).
- The iLite ® ADCC Effector (V) Assay Ready Cells (BM4001) is included as part of the iLite® Anti-CD20 ADCC Activity Set
- 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.
Product Developed and Manufactured by EuroDiagnostica
For Research Use Only
iLite® ADCC Effector (V) Assay Ready Cells are based on a human T lymphocyte cell line, Jurkat (ATCC #TIB-152), and have been genetically engineered and optimized to express high levels of the low affinity Fc receptor FcγIIIa (CD16), and the Firefly Luciferase (FL) reporter gene under the control of a proprietary chimeric promoter. The FcγIIIa receptor responds to ligation of the Fc moiety of an antibody bound to the specific antigen on target cells by activation of the FL reporter gene.
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.
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 .
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.
The Eagle Bio iLite Assay Ready Cells are designed for the specific detection of drug potency in serum/plasma as well as neutralizing antibodies. They utilize an assay technique called reporter gene assay ( RPG ) to analyze serum/plasma samples. The reporter genes utlized are encoded with a bioluminescent luciferase (Firefly luciferase). Different levels of luminescence detected from each reporter gene cell indicates different levels of expression.
Key benefits of iLite® ADCC Activity Assays:
- Unparalleled sensitivity
- High serum tolerance
- Normalization read-out included
- Negative control available for screening of unspecific activity
- Easy to use – no culturing required on target or effector cells