Fc Effector Activity Cellular Bioassays

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. 

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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 effector responses by IgG subclass/variant
Fcγ receptorFunctionIgG1IgG1NQIgG1futIgG2IgG3IgG4 (S228P)
FcγRI (CD64)ADCP+++++--++
FcγRIIa (CD32)ADCP+-+++++
FcγRIIIa (CD16) V158ADCC++-+++/-+++
FcγRIIIa (CD16) 158F lowADCC++-+++/-+++