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A Lambda-evo (λevo) phage platform for Zika virus EDIII protein display.

Honorio Negrete-Méndez1, Guadalupe Valencia-Toxqui2, Eva Martínez-Peñafiel1

  • 1Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Av. Instituto Politécnico Nacional No, 2508, C.P. 07360, Mexico City, Mexico.

Applied Microbiology and Biotechnology
|January 17, 2025
PubMed
Summary
This summary is machine-generated.

A new lambda phage (λevo) platform was developed for displaying Zika virus peptides. This phage display technology successfully generated antibodies against Zika virus in mice, showcasing its potential for vaccine development.

Keywords:
Directed evolutionLambda phageLambda-evo phagePhage displayRecombinant-proteinZIKVλevo

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Virology

Background:

  • Phage display is a significant technology for exhibiting heterologous peptides on virion surfaces.
  • Lambda phage (λ) systems offer a robust platform for displaying engineered proteins.

Purpose of the Study:

  • To develop an in vitro evolved lambda phage (λevo) platform for displaying recombinant peptides, specifically targeting Zika virus epitopes.
  • To assess the stability and display efficiency of fusion proteins on the λevo phage surface.
  • To evaluate the immunogenicity of displayed peptides in a mouse model.

Main Methods:

  • Directed evolution of lambda phage (λevo) by in vitro adaptation using Escherichia coli.
  • Construction of fusion proteins: Dλ linked to Zika virus E protein domain III (Dλ-ZE DIII) or GFP (Dλ-GFP).
  • Immunization of BALB/c mice with decorated λevo phages and subsequent antibody detection via dot blot and Western blot.

Main Results:

  • The λevo phage evolved a genome deletion, enhancing stability and enabling display of Dλ-ZE DIII and Dλ-GFP fusion proteins.
  • Despite successful decoration, Dλ-ZE DIII aggregation led to a 1000-fold reduction in viral particle production compared to wild-type.
  • Inoculation of BALB/c mice with decorated phages elicited murine antibodies against Zika virus.

Conclusions:

  • The λevo phage display platform is a versatile tool for presenting recombinant peptides, including viral antigens.
  • The platform demonstrates potential for generating immune responses against specific pathogens, like Zika virus.
  • Further optimization for protein solubility and yield is recommended for broader applications.