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Quantifying Human Norovirus Virus-like Particles Binding to Commensal Bacteria Using Flow Cytometry
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Quantifying Human Norovirus Virus-like Particles Binding to Commensal Bacteria Using Flow Cytometry

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Recombinant norovirus-specific scFv inhibit virus-like particle binding to cellular ligands

Khalil Ettayebi1, Michele E Hardy

  • 1Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717, USA. kettayeb@montana.edu

Virology Journal
|February 2, 2008
PubMed
Abstract

Insights

Engineered norovirus-binding antibody fragments (scFv54.6) effectively block virus-like particle (VLP) attachment to cells. These single chain variable fragments (scFv) show promise as a novel strategy to prevent norovirus spread and outbreaks.

Area of Science:

  • Virology
  • Immunology
  • Biotechnology

Background:

  • Noroviruses are a leading cause of epidemic gastrointestinal illness across all age groups.
  • Effective control strategies are needed due to rapid transmission and ease of person-to-person spread.
  • Inhibiting initial virus-cell binding is a promising approach to limit norovirus outbreaks.

Purpose of the Study:

  • To engineer and evaluate single chain variable fragments (scFv) derived from a norovirus-blocking monoclonal antibody (mAb 54.6).
  • To determine if scFv54.6 retains the cell-binding inhibitory functions of the parent mAb.
  • To assess the potential of scFv54.6 as a prophylactic agent against norovirus infection.

Main Methods:

  • Engineered antigen-binding domains (VL and VH) of mAb 54.6 into a single chain variable fragment (scFv54.6) with a peptide linker.
  • Expressed and purified recombinant scFv54.6 protein using Pichia pastoris.
  • Assessed scFv54.6 binding to norovirus-like particles (VLPs) and its ability to inhibit VLP-mediated hemagglutination and cell binding to H carbohydrate antigen.

Main Results:

  • Purified scFv54.6 recognized native VLPs in immunoblots.
  • scFv54.6 demonstrated inhibition of VLP-mediated hemagglutination.
  • scFv54.6 successfully blocked VLP binding to H carbohydrate antigen on transfected CHO cells.

Conclusions:

  • The engineered scFv54.6 retained the functional properties of the parent mAb 54.6.
  • scFv54.6 effectively inhibits norovirus particle interactions with host cells.
  • Norovirus-neutralizing antibodies, further engineered for mucosal delivery, offer a potential prophylactic strategy for outbreak settings.

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