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


ABSTRACT: 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 ~100 nm 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.

SUBMITTER: Pan YS 

PROVIDER: S-EPMC3004410 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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

Pan Yi-Shin YS   Wei Hung-Ju HJ   Chang Chung-Chieh CC   Lin Chung-Hung CH   Wei Ting-Shyang TS   Wu Suh-Chin SC   Chang Ding-Kwo DK  

Journal of biomedicine & biotechnology 20101209


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 ~100 nm 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 struc  ...[more]

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