Determining ADCC Activity of Antibody-Based Therapeutic Molecules using Two Bioluminescent Reporter-Based Bioassays

Area of Science:

• Biotechnology
• Immunology
• Pharmacology

Background:

• Antibody Fc effector function is a key mechanism of action for therapeutic monoclonal antibodies.
• Measuring antibody-dependent cellular cytotoxicity (ADCC) is vital for antibody development.

Purpose of the Study:

• To describe two quantitative cell-based bioassays for measuring antibody potency in ADCC.
• To provide protocols for ADCC reporter bioassay and PBMC ADCC bioassay.

Main Methods:

• Protocol 1: ADCC reporter bioassay using engineered effector cells and measuring luciferase reporter activation.
• Protocol 2: PBMC ADCC bioassay using primary PBMCs and HiBiT target cells, measuring HiBiT release upon target cell lysis.

Main Results:

• Two distinct bioassays provide quantitative measurement of antibody potency in ADCC.
• Discussion on optimizing key assay parameters like cell handling and effector:target ratios impacts assay performance.

Conclusions:

• These validated bioassays are essential tools for assessing Fc effector function in monoclonal antibody development.
• Optimization of assay parameters ensures reliable and accurate measurement of antibody potency.