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

You might also read

Related Articles

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

Sort by
Same author

Direct visualization of native GSDMD pores reveals lipid-driven stabilization during pyroptosis.

Science advances·2026
Same author

Promiscuous RNA binding by WDR5 remodels the KMT2A (MLL1) histone methyltransferase complex to an inactive state.

bioRxiv : the preprint server for biology·2026
Same author

Protein Engineering-Enabled Cryo-EM Investigation of Small GTPases.

Journal of molecular biology·2026
Same author

Structural rewiring of IL-7R dimerization by an oncogenic transmembrane mutation can be reversed by rational design.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

A Modular Toolkit for Nanoscale Interrogation of Multiprotein Assemblies Inside Living Cells.

ACS nano·2026
Same author

Hour-scale single-molecule imaging reveals dynamic assembly of the Wnt co-receptors LRP6 and ROR2 into common signalosomes.

Science signaling·2026

Related Experiment Video

Updated: Jun 20, 2026

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes
10:43

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Published on: October 3, 2019

Accelerating phage-display library selection by reversible and site-specific biotinylation.

Akiko Koide1, John Wojcik, Ryan N Gilbreth

  • 1Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA.

Protein Engineering, Design & Selection : PEDS
|September 10, 2009
PubMed
Summary

This study introduces a new method for immobilizing target molecules using His-tags and (BT)tris-NTA for enhanced phage display library sorting. This approach improves target preparation and reveals previously masked epitopes, leading to better binding protein selection.

More Related Videos

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
10:17

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
06:30

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

Published on: August 27, 2021

Related Experiment Videos

Last Updated: Jun 20, 2026

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes
10:43

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Published on: October 3, 2019

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
10:17

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
06:30

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

Published on: August 27, 2021

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Engineering

Background:

  • Phage display library sorting requires target molecule immobilization to a solid support.
  • Existing immobilization strategies have limitations that can hinder effective sorting.

Purpose of the Study:

  • To develop an improved immobilization method for phage display library sorting.
  • To simplify target preparation and enhance the selection of high-affinity binding proteins.

Main Methods:

  • Utilized a polyhistidine-tagged (His-tagged) target molecule.
  • Employed (BT)tris-NTA, a high-affinity capture reagent for His-tags with a biotin moiety.
  • Linked His-tagged molecules to a streptavidin-coated solid support via (BT)tris-NTA for stable and reversible immobilization.

Main Results:

  • Successfully selected high-affinity binding proteins using fibronectin type III (FN3) scaffold against His-tagged targets (yeast small ubiquitin-like modifier and maltose-binding protein).
  • FN3 clones selected with the new method showed significantly stronger binding compared to those selected using passive adsorption on polystyrene.
  • His-tag-mediated immobilization exposed epitopes masked by traditional adsorption methods.

Conclusions:

  • The developed His-tag-mediated immobilization method streamlines target preparation for phage display.
  • This technique enhances the discovery of binding proteins by improving epitope accessibility.
  • The method has the potential to significantly improve binding-protein engineering experiments.