iLite® ADCC Target HER2 (-) Assay Ready Cells are based on a human embryonic kidney cell line, HEK293* (ATCC# CRL-1573), and have been genetically engineered and optimized to be depleted of HER2 expression. The cells are to be use as negative controls together with iLite® ADCC Effector (V) Assay Ready Cells and iLite® ADCC Target HER2 (+) Assay Ready Cells for measuring the ADCC activity of anti-HER2 antibodies.
* The HEK-293 cell line has been used under a license obtained from AdVec Inc.
The iLite® ADCC Target HER2 (-) Assay Ready Cells can be used together with iLite® ADCC Effector (V) and iLite® ADCC Target HER2 (+) Assay Ready Cells for the quantification ADCC activity.
Application notes for the following assays are available:
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.
Breast cancer is the most common cancer in women worldwide, and the second most common cancer overall. Survival rates have improved in the recent years, and stratification of patients into subgroups has vastly improved treatment options for many patients. As an example, patients with HER2 positive breast cancer generally have a poor prognosis, but treatment with trastuzumab, a monoclonal antibody targeting the HER2 receptor, has shown to increase both overall survival and disease free survival when given together with chemotherapy. Trastuzumab’s mechanism of action is mediated in part by inducing ADCC when crosslinking HER2 positive cells with the patient's immune cells. The drug was FDA approved for treatment of breast cancer patients in 1998, and many biosimilars are currently in development.
>250 µL of Assay Ready Cells diluted in DMEM with 20% heat inactivated fetal bovine serum (FBS), mixed 1:1 with cryoprotective medium from Lonza (Cat. No 12-132A).