A Lambda-evo (λ_evo) phage platform for Zika virus E_DIII protein display

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.