Construction and Characterization of Insect Cell-Derived Influenza VLP: Cell Binding, Fusion, and EGFP Incorporation
| Autor: | Ting-Shyang Wei, Chung-Chieh Chang, Ding-Kwo Chang, Chung-Hung Lin, Suh-Chin Wu, Hung-Ju Wei, Yi-Shin Pan |
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| Rok vydání: | 2010 |
| Předmět: |
Fluorescence-lifetime imaging microscopy
Article Subject lcsh:Biotechnology viruses Health Toxicology and Mutagenesis Green Fluorescent Proteins lcsh:Medicine Neuraminidase Hemagglutinin (influenza) Sf9 Spodoptera complex mixtures Cell Line Green fluorescent protein Jurkat Cells Viral Envelope Proteins lcsh:TP248.13-248.65 Genetics Animals Humans Cloning Molecular Surface plasmon resonance Molecular Biology Viral matrix protein biology lcsh:R Virion virus diseases General Medicine Surface Plasmon Resonance Virus Internalization Molecular biology Endocytosis Recombinant Proteins Microscopy Fluorescence Cell culture biology.protein Biophysics Molecular Medicine Baculoviridae Research Article Biotechnology |
| Zdroj: | Journal of Biomedicine and Biotechnology Journal of Biomedicine and Biotechnology, Vol 2010 (2010) |
| ISSN: | 1110-7251 1110-7243 |
| DOI: | 10.1155/2010/506363 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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