Analytical approaches for the evaluation of ADCC activity of mAbs

Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) is a key mechanism of action for therapeutic monoclonal antibodies, particularly in oncology, where it enables targeted destruction of infected or cancer cells by immune effector cells. As ADCC often reflects the in vivo activity of these biopharmaceuticals, robust analytical methods are required for quality control and product characterisation.

This application note presents a validated reporter gene bioassay to assess ADCC activity of anti-CD20 antibodies, alongside complementary analytical approaches including Surface Plasmon Resonance (SPR) and high-resolution mass spectrometry for glycosylation profiling. Together, these methods provide a comprehensive evaluation of antibody functionality and critical quality attributes. The full application note is available for download below.

KEY HIGHLIGHT OF THE BIOASSAY METHOD

• Use of a reporter-gene ADCC bioassay with engineered effector cells, enabling reduced variability compared to primary cell-based assays
• Validated method demonstrating accuracy within the 50%–150% potency range, with relative bias between -9% and -1%
• Intermediate precision maintained below 20% for most levels and 18% overall
• Demonstrated linearity across a 50%–150% range with regression parameters consistent with validation requirements
• Specific response to anti-CD20 antibodies, confirming assay selectivity and reliability

SUPPORTING YOUR ADCC STRATEGY

The combined analytical approaches described in this application note support robust characterisation and quality control of monoclonal antibodies by linking functional activity with structural attributes.

• Provides a validated and reproducible bioassay for ADCC activity assessment
• Enables reduction of variability through the use of standardised, thaw-and-use effector cells
• Supports deeper understanding of structure–function relationships via correlation between FcγRIIIa binding affinity and ADCC activity
• Integrates orthogonal methods (SPR and HRMS) to strengthen analytical comparability and data interpretation
• Facilitates monitoring of critical quality attributes such as Fc glycosylation, known to impact ADCC activity