Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Recombinant DNA01:09

Recombinant DNA

99.2K
Overview
99.2K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Characterization of Fluorophore-Quencher Pairs for Distance-Dependent Molecular Sensing in Photoacoustic Imaging.

Chemical & biomedical imaging·2026
Same author

Bioinformatic pipeline to identify potential therapeutic targets with subsequent isolation and characterization of novel human anti- DDR1 antibodies.

Scientific reports·2026
Same author

Development of a high-affinity anti-ROR1 variable region for broad anti-cancer immunotherapy.

Molecular therapy : the journal of the American Society of Gene Therapy·2025
Same author

A novel chimeric coronavirus spike vaccine combining SARS-CoV-2 RBD and scaffold domains from HKU-1 elicits potent neutralising antibody responses.

NPJ vaccines·2025
Same author

Safety and Immunogenicity of a Severe Acute Respiratory Syndrome Coronavirus 2 Spike Subunit Vaccine Stabilized in the Prefusion Conformation by a Second-Generation Molecular Clamp and Evaluated in Adults Aged 18-55 Years: A Randomized, Double-Blind, Active Comparator, Phase I Trial.

The Journal of infectious diseases·2025
Same author

Automated analysis of pore structures in biomaterials.

Journal of materials chemistry. B·2025

Related Experiment Video

Updated: Nov 20, 2025

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
15:27

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

Published on: April 17, 2017

21.2K

Recombinant Antibody Engineering Enables Reversible Binding for Continuous Protein Biosensing.

Christian Fercher1,2, Martina L Jones1, Stephen M Mahler1

  • 1Australian Institute for Bioengineering and Nanotechnology, ARC Training Centre for Biopharmaceutical Innovation, The University of Queensland, St. Lucia, Queensland, 4072 Australia.

ACS Sensors
|January 22, 2021
PubMed
Summary

Directed evolution engineered a faster-dissociating antibody fragment for continuous, label-free biosensing of epidermal growth factor receptor (EGFR). This advance enables sensitive protein monitoring in complex samples.

Keywords:
antibody engineeringcontinuous biosensinglabel-freemutagenesisunnatural amino acid

More Related Videos

The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
08:06

The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions

Published on: February 1, 2018

9.3K
Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
14:43

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications

Published on: September 23, 2013

11.0K

Related Experiment Videos

Last Updated: Nov 20, 2025

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
15:27

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

Published on: April 17, 2017

21.2K
The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
08:06

The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions

Published on: February 1, 2018

9.3K
Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
14:43

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications

Published on: September 23, 2013

11.0K

Area of Science:

  • Biosensor development
  • Protein engineering
  • Antibody engineering

Background:

  • Continuous protein monitoring in complex biological samples is challenging for current affinity biosensors.
  • Engineering antibodies is crucial for improving biosensor specificity, detection limits, and functionality.

Purpose of the Study:

  • To modulate the dissociation kinetics of an anti-epidermal growth factor receptor (EGFR) single-chain variable antibody fragment (scFv) using directed evolution.
  • To enable continuous, label-free protein sensing in complex biological samples.

Main Methods:

  • Generated a mutant scFv library by permuting residues in the antibody-antigen-binding interface.
  • Utilized phage display biopanning and high-throughput screening to isolate binders with fast dissociation kinetics.
  • Demonstrated continuous sensing in a modified label-free binding assay and confirmed with fluorescence-based detection.

Main Results:

  • Isolated a mutant scFv with approximately 30 times faster dissociation rates than the wild type (WT).
  • Maintained overall affinity and specificity while significantly enhancing dissociation kinetics.
  • Achieved low nanomolar detection limits (KD = 8.39 nM) for EGFR in continuous mode.

Conclusions:

  • Directed evolution is a viable strategy to optimize antibody fragments for enhanced biosensing applications.
  • The developed mutant scFv enables sensitive and continuous monitoring of EGFR in label-free assays.
  • This generic approach can be applied to engineer binders for various proteins and biosensor platforms.