title: Adenovirus Dodecahedron, a VLP, Can be Purified by Size Exclusion Chromatography Instead of Time-Consuming Sucrose Density Gradient Centrifugation.
abstract: Adenoviral dodecahedron (Dd) is a virus-like particle composed of twelve pentameric penton base (Pb) proteins, responsible for adenovirus cell penetration. It is generated spontaneously in the baculovirus system upon expression of the Pb gene of adenovirus serotype 3. This particle shows remarkable cell penetration ability with 2,00,000-3,00,000 Dd internalized into one cell in culture, conceivably delivering several millions of foreign cargo molecules to the target cell. We have used it in the past for delivery of small drugs as well as a vaccination platform, in which Dd serves as a particulate vaccine delivery system. Since development of new biomedicals depends strongly on the cost of their expression and purification, we attempted, albeit unsuccessfully, to obtain Dd expression in bacteria. We therefore retained its expression in the baculovirus/insect cells system but introduced significant improvements in the protocols for Dd expression and purification, leading to considerable savings in time and improved yield.
authors.affiliation: Institute of Biochemistry and Biophysics of Polish Academy of Sciences, Pawińskiego 5a, 02106, Warsaw, Poland.
authors.forename: I
authors.initials: I
authors.lastname: Szurgot
authors.forename: M
authors.initials: M
authors.lastname: Jedynak
authors.forename: M
authors.initials: M
authors.lastname: Podsiadla-Bialoskorska
authors.forename: Jan
authors.initials: J
authors.lastname: Piwowarski
authors.forename: E
authors.initials: E
authors.lastname: Szolajska
authors.forename: J
authors.initials: J
authors.lastname: Chroboczek
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D002498:Centrifugation / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D013395:Sucrose; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D014764:Viral Proteins; D013395:Sucrose