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Protein Dynamics in Living Cells

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Related Experiment Video

Updated: Mar 29, 2026

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies

Published on: November 18, 2022

Real-Time Probing of Molecular Affinity Using Optical Tweezers.

Joana Teixeira^1,2, José A Ribeiro³, Marcus Monteiro³

¹Center for Applied Photonics, INESC TEC, Rua do Campo Alegre 687, 4169-007 Porto, Portugal.

Sensors (Basel, Switzerland)

|March 28, 2026

Summary

This summary is machine-generated.

This study introduces a novel optical tweezer method to track molecular binding in real time. The technique accurately measures binding kinetics for high-affinity interactions, offering a label-free approach for biochemical analysis.

Keywords:

kinetic analysis molecular affinity optical trapping signal processing

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Area of Science:

Biophysics
Biochemistry
Nanotechnology

Background:

Real-time assessment of molecular binding kinetics is crucial for understanding biochemical and biotechnological processes.
Existing methods for monitoring molecular interactions have limitations in real-time, label-free detection and adaptability.

Purpose of the Study:

To present a novel optical tweezer (OT)-based method for real-time monitoring of molecular affinity.
To demonstrate the method's capability using the high-affinity streptavidin-biotin system as a model.

Main Methods:

Utilized transparent poly(methyl methacrylate) (PMMA) microparticles functionalized with streptavidin.
Trapped microparticles and analyzed forward-scattered signals to detect nanoscale size changes upon binding with biotinylated bovine serum albumin (biotin-BSA).
Applied the Power Spectral Density method to calculate friction coefficients and track binding dynamics.

Main Results:

Successfully monitored molecular binding in real time.
Estimated the association rate constant (kon≈10^6 M^-1s^-1) for the streptavidin-biotin interaction, consistent with literature values.
Demonstrated nanoscale changes in particle size indicative of molecular binding.

Conclusions:

The developed OT-based method provides a non-invasive, label-free approach for real-time detection of molecular interactions.
This technique offers advantages over existing methods, including real-time monitoring and adaptability to various bioaffinity systems.
Establishes a foundation for using OT tools to monitor high-affinity molecular interactions, with potential for miniaturized applications.