A Method for High-Throughput Screening of Monoclonal Antibody Internalization Using a DNA/Protein Molecular Staple

Area of Science:

• Immunology
• Biotechnology
• Cell Biology

Background:

• Monoclonal antibodies (mAbs) are crucial therapeutics, but their efficacy depends on understanding interactions with cell receptors.
• Assessing mAb binding and subsequent cellular internalization is vital for therapeutic development.
• Current methods for measuring internalization are challenging due to difficulties in functionalizing and purifying mAbs with sensors.

Purpose of the Study:

• To develop a simple, one-pot method for quantifying mAb binding and internalization without chemical mAb modification.
• To enable rapid screening of mAb-receptor interactions and internalization efficiencies.
• To investigate the relationship between mAb binding affinity and cellular internalization.

Main Methods:

• Developed a protein staple to link a DNA-based internalization sensor to mAbs.
• Utilized the sensor to quantify mAb binding and internalization into T cell and B cell receptors on human peripheral blood mononuclear cells.
• Screened various lymphocyte receptors to assess binding and internalization differences.

Main Results:

• Demonstrated a novel, one-pot approach requiring no chemical mAb modification for efficient quantification of binding and internalization.
• Showed that high mAb binding to cell receptors does not necessarily correlate with high cellular uptake or internalization efficiency.
• Identified that receptor internalization efficiency is influenced by intrinsic properties of both the receptor and the mAb.

Conclusions:

• The developed method provides a simplified way to assess critical mAb functions for therapeutic development.
• High binding affinity is not a sole predictor of mAb efficacy; internalization efficiency must also be considered.
• Screening for both binding and internalization is essential for optimizing mAb therapeutics and targeted delivery systems.