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

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.