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Construction and characterization of insect cell-derived influenza VLP: Cell binding, fusion, and EGFP incorporation

Journal of Biomedicine and Biotechnology, ISSN: 1110-7243, Vol: 2010, Page: 506363
2010
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Article Description

We have constructed virus-like particles (VLPs) harboring hemagglutinin (HA), neuraminidase (NA), matrix protein 1 (M1), and proton channel protein (M2) using baculovirus as a vector in the SF9 insect cell. The size of the expressed VLP was estimated to be ∼100nm by light scattering experiment and transmission electron microscopy. Recognition of HA on the VLP surface by the HA2-specific monoclonal antibody IIF4 at acidic pH, as probed by surface plasmon resonance, indicated the pH-induced structural rearrangement of HA. Uptake of the particle by A549 mediated by HA-sialylose receptor interaction was visualized by the fluorescent-labeled VLP. The HA-promoted cell-virus fusion activity was illustrated by fluorescence imaging on the Jurkat cells incubated with rhodamine-loaded VLP performed at fusogenic pH. Furthermore, the green fluorescence protein (GFP) was fused to NA to produce VLP with a pH-sensitive probe, expanding the use of VLP as an antigen carrier and a tool for viral tracking. Copyright © 2010 Yi-Shin Pan et al.

Bibliographic Details

Pan, Yi-Shin; Wei, Hung-Ju; Chang, Chung-Chieh; Lin, Chung-Hung; Wei, Ting-Shyang; Wu, Suh-Chin; Chang, Ding-Kwo

Hindawi Limited

Biochemistry, Genetics and Molecular Biology; Environmental Science

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