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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

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Published on: April 17, 2017

Continuous Protein Sensing Using Fast-Dissociating Antibody Fragments in Competition-Based Biosensing by Particle

Claire M S Michielsen^1,2, Yu-Ting Lin³, Junhong Yan³

¹Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5612 AE, the Netherlands.

|March 24, 2025

Summary

This summary is machine-generated.

This study introduces a new continuous protein monitoring method using tethered particle motion (t-BPM) with fast-dissociating antibody fragments (Fabs). The developed sensor achieved nanomolar detection of lactoferrin in various matrices for extended periods.

Keywords:

antibody fragments binding kinetics biosensing by particle motion competition biosensor continuous protein sensing

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Area of Science:

Biotechnology
Biosensing
Immunology

Background:

Continuous monitoring of protein concentrations is crucial for biological systems and bioprocess control.
Existing sensing technologies may have limitations in real-time protein quantification.

Purpose of the Study:

To develop and demonstrate a continuous protein sensing methodology using tethered particle motion (t-BPM).
To utilize fast-dissociating antibody fragments (Fabs) for enhanced protein detection.
To validate the sensor's performance for lactoferrin detection in diverse matrices.

Main Methods:

Development of a competition-based t-BPM sensor.
Characterization of custom-made Fabs using free particle motion sensing and surface plasmon resonance.
Integration of selected Fabs into the t-BPM sensor for continuous monitoring.

Main Results:

Six out of thirteen Fabs exhibited rapid association and dissociation kinetics.
The t-BPM sensor successfully detected lactoferrin at nanomolar concentrations.
Continuous detection was achieved in both buffer solutions and milk matrices over tens of hours.

Conclusions:

Fast-dissociating Fabs can be effectively employed in a competitive t-BPM sensor format for continuous protein monitoring.
This methodology offers a promising approach for real-time protein quantification in complex biological samples.