title: Enhanced gene delivery by avidin-displaying baculovirus.
abstract: Flexible alteration of virus surface properties would be beneficial for enhanced and targeted gene delivery. A useful approach could be based on a high-affinity receptor-ligand pair, such as avidin and biotin. In this study, we have constructed an avidin-displaying baculovirus, Baavi. Avidin display was expected to enhance cell transduction due to the high positive charge of avidin in physiological pH and to provide a binding site for covering the virus with desired biotinylated ligands. Successful incorporation of avidin on the virus envelope was detected by immunoblotting and electron microscopy. Multiple biotin-binding sites per virus were detected with fluorescence-correlation spectroscopy and tight biotin binding was observed using an optical biosensor, IAsys. Baavi showed a 5-fold increase in transduction efficiency in rat malignant glioma cells (BT4C) and a 26-fold increase in rabbit aortic smooth muscle (RAASMC) cells compared to wild-type baculovirus. Enhanced transduction was also observed with biotinylated target cells. Biotinylated epidermal growth factor (EGF) enabled specific targeting of the virus with high efficiency to EGF receptor-expressing (SKOV-3) cells. An additional advantage of the avidin display was demonstrated with biotinylated paramagnetic particles, which enabled magnetic targeting. Altogether, we show that avidin display is a rapid and versatile method to improve viral properties for gene delivery.
authors.affiliation: A. I. Virtanen Institute, Department of Biotechnology and Molecular Medicine, University of Kuopio, P.O. Box 1627, FIN-70120 Kuopio, Finland.
authors.forename: Jani K
authors.initials: JK
authors.lastname: Räty
authors.forename: Kari J
authors.initials: KJ
authors.lastname: Airenne
authors.forename: Ari T
authors.initials: AT
authors.lastname: Marttila
authors.forename: Varpu
authors.initials: V
authors.lastname: Marjomäki
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Hytönen
authors.forename: Pauliina
authors.initials: P
authors.lastname: Lehtolainen
authors.forename: Olli H
authors.initials: OH
authors.lastname: Laitinen
authors.forename: Anssi J
authors.initials: AJ
authors.lastname: Mähönen
authors.forename: Markku S
authors.initials: MS
authors.lastname: Kulomaa
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
mesh_terms: D000818:Animals; D001360:Avidin / Q000235:genetics / Q000378:metabolism*; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology*; D015374:Biosensing Techniques; D001710:Biotin / Q000378:metabolism; D019921:Biotinylation; D002460:Cell Line; D045744:Cell Line, Tumor; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D013050:Spectrometry, Fluorescence; D014161:Transduction, Genetic; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D014760:Viral Fusion Proteins; C030473:biotinyl epidermal growth factor; D001360:Avidin; D004815:Epidermal Growth Factor; D001710:Biotin; D066246:ErbB Receptors