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A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
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Phage display for generating peptide reagents.

Jennifer R Brigati1, Tatiana I Samoylova, Prashanth K Jayanna

  • 1Division of Natural Sciences, Maryville College, Maryville, Tennessee, USA.

Current Protocols in Protein Science
|April 23, 2008
PubMed
Summary

Researchers detail methods for creating landscape phages, which display foreign peptides on their surface. These engineered phages specifically target various cells, spores, and molecular targets for potential applications.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Virology

Background:

  • Filamentous phages like fd, M13, and f1 are well-characterized viral systems.
  • Displaying foreign peptides on phage surfaces enables novel applications in molecular targeting and diagnostics.

Purpose of the Study:

  • To provide detailed protocols for the selection and propagation of landscape phages.
  • To enable the creation of chimeric phage particles displaying foreign peptides.

Main Methods:

  • Fusion of filamentous phage fd (or relatives) with foreign DNA.
  • Engineering chimeric phage virions to display peptides on the entire surface.
  • Selection and propagation protocols for engineered phages.

Main Results:

  • Successful generation of landscape phages with foreign peptides (8-9 amino acids) on their surface.
  • Demonstrated specific binding of landscape phages to mammalian cells, bacterial cells, spores, and molecular targets.

Conclusions:

  • Landscape phages offer a versatile platform for targeted molecular interactions.
  • The presented protocols facilitate the development of phage-based tools for diverse biological applications.