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Related Concept Videos

Genetic Screens02:46

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...

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Related Experiment Video

Updated: May 26, 2026

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
12:36

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits

Published on: February 16, 2014

Phage display screening without repetitious selection rounds.

Peter A C 't Hoen1, Silvana M G Jirka, Bradley R Ten Broeke

  • 1Center for Human and Clinical Genetics and Leiden Genome Technology Center, Leiden University Medical Center, 2300 RC Leiden, The Netherlands. p.a.c.hoen@lumc.nl

Analytical Biochemistry
|December 20, 2011
PubMed
Summary
This summary is machine-generated.

Next-generation sequencing (NGS) accelerates phage display by identifying specific binders and reducing false positives. Deep sequencing after just one biopanning round reveals true phage hits, improving peptide and antibody discovery.

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Last Updated: May 26, 2026

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Immunology

Background:

  • Phage display is a common method for identifying high-affinity peptides and antibodies.
  • However, it is labor-intensive and prone to false positive results.

Purpose of the Study:

  • To enhance phage display efficiency and accuracy using next-generation sequencing (NGS).
  • To characterize phage libraries before and after biopanning to identify sources of error.

Main Methods:

  • Illumina next-generation sequencing was used to analyze a Ph.D.-7 M13 peptide phage display library.
  • Deep sequencing was performed on the naive library and after biopanning rounds on KS483 osteoblast cells.
  • Confocal and live cell imaging validated peptide binding and uptake.

Main Results:

  • Sequencing revealed that propagation advantage in bacteria can lead to false positive hits.
  • Deep sequencing after a single biopanning round was sufficient to identify positive phages.
  • This approach avoids the loss of slower-propagating, yet promising, clones.

Conclusions:

  • NGS significantly improves phage display by speeding up the identification of specific binders.
  • Utilizing NGS reduces the number of false positive hits in phage display screenings.
  • This method enhances the discovery of peptides and antibodies for therapeutic applications.