Engineered Jurkat-NFAT Reporter cells for ADCC and ADCP
InvivoGen’s cell-based effector activity reporter assays provide biologically relevant tools to assess antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). They are designed to evaluate the potency and Fc-effector activity of monoclonal antibodies (mAbs) and other biologics that engage with Fc-gamma receptors (FcγR).
These bioluminescent assays are based on Jurkat-Lucia™ effector cells stably expressing a specific Fcγ receptor variant, either CD64 (FcγRI), CD32A (FcγRIIa), CD16A (FcγRIIIa) high-affinity allotype V158, or CD16A low-affinity allotype F158, together with an NFAT-inducible Lucia® luciferase reporter gene.
Key features
- No primary cell culture required
- Robust and reproducible assay design
- Sensitive Lucia® luciferase readout
- Suitable for antibody potency testing and Fc engineering
Applications
- Screening of potent biologics for ADCC or ADCP induction
- Evaluation of Fc engineering strategies to modulate effector potency
- Testing of novel anti-CD16, -CD32, or -CD64 mAbs
Co-culture of Jurkat-Lucia™ NFAT effector cells with antigen-presenting target cells (e.g., Raji cells) in the presence of test antibodies leads to NFAT-mediated Lucia® reporter activation. The intensity of the Lucia® signal provides a direct, quantitative measure of the antibody’s Fc-mediated effector function.
Fcγ receptor | Function | IgG1 | IgG1NQ | IgG1fut | IgG2 | IgG3 | IgG4 (S228P) |
---|---|---|---|---|---|---|---|
FcγRI (CD64) | ADCP | ++ | + | ++ | - | - | ++ |
FcγRIIa (CD32) | ADCP | + | - | + | ++ | + | + |
FcγRIIIa (CD16) V158 | ADCC | ++ | - | ++ | +/- | ++ | + |
FcγRIIIa (CD16) 158F low | ADCC | ++ | - | ++ | +/- | ++ | + |