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

One of the most significant bacteriophage technologies is phage display, in which heterologous peptides are exhibited on the virion surface. This work describes the display of λ decorative protein D_λ linked to the E protein domain III of Zika virus (D_λ-ZE_DIII), to the GFP protein (D_λ-GFP), or to different domain III epitopes of the E_ZIKV protein (D_λ-TD), exhibited on the surface of an in vitro evolved lambda phage (λ_evo). This phage harbors a gene D deletion and was subjected to directed evolution using Escherichia coli W3110/pD_λ-ZE_DIII as background. After 20 days (20 cycles of dilution), the λ_evo phage developed a ~ 22% genome deletion affecting the non-essential λ b region, rendering a more stable phage that exhibited fusion proteins D_λ-ZE_DIII or D_λ-GFP but not D_λ-TD. Despite the λ_evo system was able to decorate itself with the D_λ-ZE_DIII protein, the production of viral particles was ~ 1000-fold lower than the λ wild-type, due to the unexpected D_λ-ZE_DIII protein aggregation into bacterial inclusion bodies. Decorated phages (10⁶ PFU (plaque forming units)/100 µl) were inoculated into BALB/c mice, and subsequent dot blot and Western blot immunoassays proved the production of murine antibodies against ZIKV (Zika virus). This multipurpose λ_evo phage display platform may be used interchangeably with other more soluble peptides, providing better yields.

• λ_evo platform for displaying recombinant peptides.

• Directed evolution to generate λ_evo with more efficient decoration.

• Antigenic reaction in BALB/c mice by inoculating λ_evo with recombinant peptides.