Probing fibronectin-antibody interactions using AFM force spectroscopy and lateral force microscopy

Area of Science:

• Biophysics
• Surface Science
• Nanotechnology

Background:

• Lateral Force Microscopy (LFM) allows studying specific interaction forces, like lectin-carbohydrate binding.
• Quantitative LFM measurements depend on accurate torsional force calibration, which is complex and time-consuming.

Purpose of the Study:

• To present a novel, simplified method for calibrating torsional force in LFM.
• To validate the new calibration method using a relevant biological system.

Main Methods:

• Development of a new, simplified torsional force calibration technique for LFM.
• Application of LFM with the new calibration to measure forces between human fibronectin and its monoclonal antibody.
• Comparison of LFM results with Atomic Force Microscopy (AFM)-based force spectroscopy.

Main Results:

• The proposed method offers a simpler approach to torsional force calibration in LFM.
• Similar unbinding forces were observed using LFM and AFM at comparable loading rates.
• The study successfully measured single molecule interactions between fibronectin and its antibody.

Conclusions:

• The developed lateral force calibration method is effective and applicable for single molecule interaction studies.
• This advancement simplifies quantitative analysis in LFM experiments.
• The method provides reliable data for biophysical interaction studies.