Antibody dependent Cell Cytotoxicity (ADCC) assessment

Consisting in the targeting of cancer cells for destruction by antibodies to be then killed by effector immune cells, antibody-dependent cell cytotoxicity (ADCC) is thus the basis of several monoclonal antibody therapies which have been proven powerful and highly promising for cancer treatment.
Explicyte offers to perform an ADCC assay based on the full-time course monitoring of both cell proliferation and death of tumor cells, in the presence of immune cells and of candidate antibodies targeting specific tumor antigens. Our kinetic and quantitative cell-based assay allows thus the dynamic assessment and evaluation of the efficacy of test antibodies in eliciting ADCC and directing killing activity towards target tumor cells.

To learn more, download our antibody-dependent NK cell-mediated cytotoxicity white paper!

Assay principle


  • Co-culture of target tumor cells with effector immune cells (PBMCs, NK cells…).
  • Real-time monitoring of tumor cell count (tumor cells stably expressing a nuclear fluorescent probe) as a surrogate of cell proliferation and of tumor cell death.
  • Automated image analysis & segmentation enables apoptotic death quantitation specifically within tumor cells (caspase 3/7 fluorescent probe within the nuclear probe-expressing cells).
  • IFNgamma release (or other effector cytokines) quantification as a surrogate of functionality of effector immune cells. 

Illustrative data

NK-mediated cytotoxicity in SKOV3 ovarian tumor cells
Combination of NK cells with anti-HER2 or anti-EGFR antibodies enhances NK-mediated cytotoxicity in SKOV3 ovarian tumor cells, in a NK ratio-dependent manner.
SKOV3 tumor cells were cultured under control, Cetuximab-, or Trastuzumab-treated conditions and, 24h later, IL2- activated primary NK cells were added at different E:T ratios. 24h later, supernatants were collected and analyzed for IFN𝜸 release by HTRF as a surrogate of NK-induced cell toxicity.
While IFN𝜸 levels are increased in a NK-ratio dependent manner, Cetuximab and Trastuzumab treatments of tumor cells further promote this increase whatever the E:T ratio, thereby showing an ADCC activity.

Combination of NK cells with anti-HER2 or anti-EGFR antibodies enhances NK-mediated cytotoxicity ...

Anti-EGFR antibody-dependent cell cytotoxicity (ADCC)
Dose-dependent anti-EGFR antibody (Cetuximab)-dependent cell cytotoxicity in SKOV3 ovarian tumor cells.
SKOV3 tumor cells were cultured under control or increasing doses Cetuximab-treated conditions and, 24h later, IL2- activated primary NK cells were added at an appropriate E:T ratio (3:1).
(A) Integration of live cell imaging data at 15h after NK cell addition as AUC of tumor cell apoptosis as a surrogate measure of NK-mediated tumor cytotoxicity.
(B) 24h following NK cell addition, supernatants were collected and analyzed for IFN𝜸 release by HTRF.

Dose-dependent anti-EGFR antibody (Cetuximab)-dependent cell cytotoxicity in SKOV3 ovarian ...