iCrea™ ADCC EGFR (+) Target Cell Line, Human HEK293 Cells (CAT#: AD-P015-C) Datasheet

iCrea? ADCC Target EGFR (+) Assay Cell Line are human cells engineered to overexpress EGFR and optimized to give high sensitivity and specificity when used together with iLite ADCC Effector (V) Assay Cell Line. iLite ADCC Target EGFR (+) Assay Cell Line are based on a human embryonic kidney cell line, HEK2931 ( CRL-1573), and have been genetically engineered and optimized to express high and constant levels of the surface antigen EGFR. The cells are to be used as target cells together with iCrea? ADCC Effector (V) Assay Cell Line for measuring the ADCC activity of anti-EGFR antibodies.

Read-out of results: Quantitative
Detection system: Luminescence
Regulatory status: Research Use Only

The iCrea? EGFR (+) Target Assay Cell Line can be used together with iCrea? ADCC Effector (V) and iCrea? EGFR (-) Target Assay Cell Line for the quantification ADCC activity of anti-EGFR antibodies.

The iCrea? ADCC Assay Cell Line 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 targetbound 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 (iCrea? ADCC Effector (V) Assay Cell Line) 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 EGFR (iCrea? EGFR (+) Target Assay Cell Line). iCrea? ADCC Effector (V) Assay Cell Line also contain the Renilla Luciferase (RL) 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 RL. 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 EGFR expression (iCrea? EGFR (-) Target Assay Cell Line). 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 cetuximab in the sample (Fig 1).