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- Title
Recombinant norovirus-specific scFv inhibit virus-like particle binding to cellular ligands.
- Authors
Ettayebi, Khalil; Hardy, Michele E.
- Abstract
Background: Noroviruses cause epidemic outbreaks of gastrointestinal illness in all age-groups. The rapid onset and ease of person-to-person transmission suggest that inhibitors of the initial steps of virus binding to susceptible cells have value in limiting spread and outbreak persistence. We previously generated a monoclonal antibody (mAb) 54.6 that blocks binding of recombinant norovirus-like particles (VLP) to Caco-2 intestinal cells and inhibits VLP-mediated hemagglutination. In this study, we engineered the antigen binding domains of mAb 54.6 into a single chain variable fragment (scFv) and tested whether these scFv could function as cell binding inhibitors, similar to the parent mAb. Results: The scFv54.6 construct was engineered to encode the light (VL) and heavy (VH) variable domains of mAb 54.6 separated by a flexible peptide linker, and this recombinant protein was expressed in Pichia pastoris. Purified scFv54.6 recognized native VLPs by immunoblot, inhibited VLP-mediated hemagglutination, and blocked VLP binding to H carbohydrate antigen expressed on the surface of a CHO cell line stably transfected to express α 1,2-fucosyltransferase. Conclusion: scFv54.6 retained the functional properties of the parent mAb with respect to inhibiting norovirus particle interactions with cells. With further engineering into a form deliverable to the gut mucosa, norovirus neutralizing antibodies represent a prophylactic strategy that would be valuable in outbreak settings.
- Subjects
NOROVIRUSES; RECOMBINANT viruses; VIRAL genetics; LIGANDS (Biochemistry); GASTROINTESTINAL diseases; CELL adhesion; MONOCLONAL antibodies; BLOOD agglutination
- Publication
Virology Journal, 2008, Vol 5, p1
- ISSN
1743-422X
- Publication type
Article
- DOI
10.1186/1743-422X-5-21