iLite anti-HER2 ADCC Activity Set

iLite anti-HER2 ADCC Activity Set Developed and Manufactured by Svar Life Science

The Eagle Biosciences iLite® anti-HER2 ADCC Activity Set includes (components are also sold separately):

• iLite® ADCC Effector (V) Assay Ready Cells (BM5001)
• iLite® ADCC Target HER2 (+) Assay Ready Cells (BM5011)
• iLite® ADCC Target HER2 (-) Assay Ready Cells (BM5016)

For Research Use Only

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

Assay Principle

The iLite® ADCC Assay Ready Cells are engineered cells that enable antibody-dependent cell-mediated cytotoxicity (ADCC) to be examined through the specific expression of Firefly luciferase. When the antibodies of interest bind to the antigens on the surface of the target cell, the target-bound antibodies will be presented to the Fc receptors (FcγRIIIa) on the effector cell. When the Fc-portion of the target-bound antibodies binds to the receptor, multiple cross-linking of the two cell types occurs. This will initiate a signaling cascade which triggers the expression of Firefly luciferase (FL) in the effector cell. In this application note, we describe the use of an effector cell line (iLite® ADCC Effector (V) Assay Ready Cells) that over-express FcγRIIIa and contain the FL reporter gene that responds to the principal transcription factors that mediate signaling from the FcγRIIIa receptor, together with a positive target cell line which over-expresses the surface antigen HER2 (iLite® ADCC Target HER2 (+) Assay Ready Cells). iLite® ADCC Effector (V) Assay Ready Cells also contain the Nano Luciferase (NL) reporter gene, under the control of a constitutive promoter, that allows drug-induced FL activity to be normalized with respect to the constitutive expression of NL. This renders assay results independent of variations in cell number, serum matrix effects, or lysis of the effector cells by the target cells. In addition, we also describe the use of a negative control in the form of a target cell line depleted of HER2 expression (iLite® Target HER2 (-) Assay Ready Cells). The Firefly luciferase signal can be measured in a luminometer following addition and incubation of luciferase substrate. The Firefly luciferase signal is proportional to the functional activity of trastuzumab in the sample.