Antibody-Dependent Cell-mediated Cytotoxicity (ADCC) assay

Explicyte offers to perform an ADCC assay to monitor in real time both cell death and cell proliferation of cancer cells in the presence of immune effector cells and of candidate antibodies targeting specific tumor antigens. This assay allows the dynamic assessment of antibodies capacity to induce ADCC instead of relying on single endpoint measurements, thus improving the detection of even subtle changes in the responses.

Principle of Antibody dependent mediated cell cytotoxicity live Assay

Assay Principle

 

  • Target tumor cells are co-cultured with the immune cells of choice (e.g., purified T cells or PBMCs, NK cell line etc..).
  • Tumor cell count is enabled by labeling target cells with a range of NucLightTM Live-Cell Labeling Reagents. Tumor cell death is measured directly and in real-time by adding the mix-and-read IncuCyteTM Caspase 3/7 Reagent to the cultures.
  • Automated image analysis enables selective quantitation of tumor cell count and cell death in the continuous presence of lymphocyte populations and without separation or liquid sampling.

Key Advantages

Live-cell-ADCC-assay-comparison-table-2

Representative Results

ADCC: Trastuzumab (Herceptin®) induced immune cell killing of SKOV-3 ovarian cancer cells.  Time courses of trastuzumab-induced concentration-dependent inhibition of cell proliferation in SKOV-3 tumour cells (Her2-positive) co-cultured with PBMCs. SKOV-3 tumor cells were identified and counted by using a fluorescent marker (nuc NucLight™ Red) Concentration-response curve to trastuzumab for inhibition of proliferation in SKOV-3 cells (B). 

ADCC: Trastuzumab (Herceptin®) induced immune cell killing of SKOV-3 ovarian cancer cells.  Time courses of trastuzumab-induced concentration-dependent inhibition of cell proliferation in SKOV-3 tumour cells (Her2-positive) co-cultured with PBMCs. SKOV-3 tumor cells were identified and counted using the IncuCyte® live-cell analysis system fluorescent marker (NucLight™ Red) Concentration-response curve to trastuzumab for inhibition of proliferation in SKOV-3 cells (B).

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Explicyte immuno-oncology offers to perform a kinetic ADCC cell-based assay by live-cell imaging for the identification and profiling of novel cancer immunotherapy strategies.