Principle of ADCC Assay

1. Target cells (e.g., cancer cells or virus-infected cells) express the antigen of interest.

2. Therapeutic antibody (IgG1) binds to the antigen on the target cell surface.

3. Effector cells (e.g., NK cells, monocytes, or engineered cell lines) recognize the antibody via CD16 (FcγRIII) on their surface.

4. Effector cells release cytotoxic molecules (e.g., perforin, granzymes, TNF-α) leading to target cell lysis.

5. Cytotoxicity is measured using various readouts (e.g., LDH release, luciferase, or flow cytometry).

Model Description

Cancer cells expressing the target antigen are co-cultured with NK cells  and the monoclonal antibody, resulting in target cell destruction.

Readouts

Tumor cell destruction by flow cytometry,  NK cells response by cytokine secretion (IFNγ release) and Luciferase-based reporter expression.

Add-On Services

Evaluation of cytokine release, NK cell cytotoxic factors (perforin/granzymes), proteomics, and transcriptomics.

Key Components of ADCC Assay

Target Cells

• Should express the specific antigen recognized by the monoclonal antibody (Tumor cells or virus infected cells).

Effector Cells

• Natural Killer (NK) cells (primary mediators of ADCC).

• Peripheral blood mononuclear cells (PBMCs) (contain NK cells & monocytes).

• Jurkat NFAT-Luc reporter cells expressing CD16 (engineered cell lines for FcγR activation).

Antibodies

• Monoclonal antibodies (e.g., IgG1 with strong FcγR binding).

• Engineered antibodies with enhanced Fc-mediated effector functions.

Detection Methods

• Cytokine secretion, Flow cytometry based apoptotic cell detection and Luciferase-based reporter expression.