Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

Engineering soluble monomeric streptavidin with reversible biotin binding capability.

Sau-Ching Wu¹, Sui-Lam Wong

¹Department of Biological Sciences, University of Calgary, Alberta, Canada.

The Journal of Biological Chemistry

|April 21, 2005

Summary

Researchers engineered monomeric streptavidin with reversible biotin binding. This modified protein, designed to prevent aggregation, shows promise for various biotechnological applications requiring controlled biotin interaction.

Related Concept Videos

You might also read

Related Articles

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

Sort by

Same author

An engineered redox-switchable streptavidin mutein enables the high-affinity capture and efficient elution of biotinylated ligands.

Protein science : a publication of the Protein Society·2025

Same author

Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity.

Scientific reports·2020

Same author

A bio-coupling approach using a dextran-binding domain to immobilize an engineered streptavidin to Sephadex for easy preparation of affinity matrix.

Scientific reports·2019

Same author

A simple approach for preparation of affinity matrices: Simultaneous purification and reversible immobilization of a streptavidin mutein to agarose matrix.

Scientific reports·2017

Same author

Engineering Streptavidin and a Streptavidin-Binding Peptide with Infinite Binding Affinity and Reversible Binding Capability: Purification of a Tagged Recombinant Protein to High Purity via Affinity-Driven Thiol Coupling.

PloS one·2015

Same author

Structure-guided design of an engineered streptavidin with reusability to purify streptavidin-binding peptide tagged proteins or biotinylated proteins.

PloS one·2013

Area of Science:

Biochemistry
Protein Engineering
Biotechnology

Background:

Streptavidin's biotin binding site requires inter-subunit interactions, making natural tetramers difficult to monomerize.
Monomerization of streptavidin can reduce biotin binding affinity due to exposed hydrophobic residues.

Purpose of the Study:

To engineer monomeric streptavidin with retained or enhanced reversible biotin binding capabilities.
To develop strategies for preventing aggregation in monomeric streptavidin muteins.

Main Methods:

Site-directed mutagenesis to introduce electrostatic repulsion and steric hindrance at subunit interfaces.
Engineering for improved hydrophilicity to minimize aggregation.
Confirmation of monomerization using gel filtration, dynamic light scattering, proteinase K sensitivity, and chemical cross-linking.

Related Experiment Videos

Main Results:

A quadruple mutein (T76R,V125R,V55T,L109T) demonstrated stable monomeric state above 2 mg/ml.
This mutein exhibited excellent reversible biotin binding kinetics.
A different mutein (D61A,W120K) failed to monomerize, highlighting the importance of residue selection.

Conclusions:

Engineered monomeric streptavidin with reversible biotin binding is achievable.
Strategic mutations can effectively monomerize streptavidin and prevent aggregation.
The developed mutein offers potential for applications requiring controlled biotin interaction and purification.