A Lambda-evo (λ) Phage Platform for Zika Virus E Protein Display

Overview

Journal Appl Microbiol Biotechnol

Date 2025 Jan 17

PMID 39820667

Authors

Honorio Negrete-Mendez

Guadalupe Valencia-Toxqui

Eva Martinez-Penafiel

Oscar Medina-Contreras

Fernando Fernandez-Ramirez

Edgar Morales-Rios

Luis Janiel Navarro-Gonzalez

Jesus M Torres-Flores

Luis Kameyama

Affiliations

Soon will be listed here.

Abstract

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 linked to the E protein domain III of Zika virus (D-ZE), to the GFP protein (D-GFP), or to different domain III epitopes of the E protein (D-TD), exhibited on the surface of an in vitro evolved lambda phage (λ). This phage harbors a gene D deletion and was subjected to directed evolution using Escherichia coli W3110/pD-ZE as background. After 20 days (20 cycles of dilution), the λ phage developed a ~ 22% genome deletion affecting the non-essential λ b region, rendering a more stable phage that exhibited fusion proteins D-ZE or D-GFP but not D-TD. Despite the λ system was able to decorate itself with the D-ZE protein, the production of viral particles was ~ 1000-fold lower than the λ wild-type, due to the unexpected D-ZE protein aggregation into bacterial inclusion bodies. Decorated phages (10 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 phage display platform may be used interchangeably with other more soluble peptides, providing better yields. KEY POINTS: • λ platform for displaying recombinant peptides. • Directed evolution to generate λ with more efficient decoration. • Antigenic reaction in BALB/c mice by inoculating λ with recombinant peptides.

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