title: Fluorescence molecular painting of enveloped viruses.
abstract: In this study, we describe a versatile, flexible, and quick method to label different families of enveloped viruses with glycosylphosphatidylinositol-modified green fluorescent protein, termed fluorescence molecular painting (FMP). As an example for a potential application, we investigated virus attachment by means of flow cytometry to determine if viral binding behavior may be analyzed after FMP of enveloped viruses. Virus attachment was inhibited by using either dextran sulfate or by blocking attachment sites with virus pre-treatment. Results from the FMP-flow cytometry approach were verified by immunoblotting and enzyme-linked immunosorbent assay. Since the modification strategy is applicable to a broad range of proteins and viruses, variations of this method may be useful in a range of research and applied applications from bio-distribution studies to vaccine development and targeted infection for gene delivery.
authors.affiliation: Institute of Virology, University of Veterinary Medicine, Vienna, Veterinarplatz 1, 1210 Vienna, Austria. christoph.metzner@vetmeduni.ac.at
authors.forename: Christoph
authors.initials: C
authors.lastname: Metzner
authors.forename: Feliks
authors.initials: F
authors.lastname: Kochan
authors.forename: John A
authors.initials: JA
authors.lastname: Dangerfield
mesh_terms: D000818:Animals; D016264:Dextran Sulfate / Q000378:metabolism; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D058851:GPI-Linked Proteins / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006564:Herpesviridae / Q000254:growth & development / Q000302:isolation & purification; D015151:Immunoblotting; D009980:Influenza A virus / Q000254:growth & development / Q000302:isolation & purification; D016086:Lentivirus / Q000254:growth & development / Q000302:isolation & purification; D061985:Madin Darby Canine Kidney Cells; D010802:Phylogeny; D014771:Virion / Q000378:metabolism; D053585:Virus Attachment
chemical_list: D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols; D049452:Green Fluorescent Proteins; D016264:Dextran Sulfate