NFAT-CD16 cells are engineered from the human T lymphocyte Jurkat cell line. Jurkat cells naturally express a functional NFAT (nuclear factor that activates T cells) transcription factor, which is involved in the early signaling event of antibody-dependent cellular cytotoxicity (ADCC). ADCC is an immune mechanism through which effector cells with Fc receptors can recognize and kill antibody (Ab)-coated target cells that express antigen on their surface. ADCC is triggered by cross-linking between Abs bound by immune effector cell surface antigens and the Fc receptor CD16A. These interactions induce an increase in intracellular calcium concentration, calreticulin/calmodulin-mediated dephosphorylation of NFAT, nuclear translocation and binding to the promoter region of ADCC-related genes. Eventually, the effector cells release cytotoxic particles that kill the target cells. Jurkat NFAT-CD16 cells have been designed as effector reporter cells for our ADCC analysis. These cells stably express the Lucia luciferase reporter gene under the control of ISG54 minimal promoter fused with six NFAT response elements. ADCC induction is measured by the bioluminescence signal generated by luciferase produced by adding the appropriate detection reagent Luc.

•Stable expression of cell surface Fc receptor CD16A (FcgRIIIA; V158 high-affinity allotype)
•Stable expression of Lucia luciferase reporter gene under the control of ISG54 minimal promoter fused with six NFAT response elements
•Resistance to elastin

Measure Potency and Stability of Biobetter Antibodies that Bind and Activate ADCP through CD16