title: Purification and concentration of non-infectious West Nile virus-like particles and infectious virions using a pseudo-affinity Cellufine Sulfate column.
abstract: Affinity column chromatography is a promising method for the purification of flavivirus particles that can supplement or potentially replace diafiltration and sucrose density centrifugation. In this study, the purification of West Nile Virus (WNV) antigens via Cellufine Sulfate column chromatography was examined. Virus-like particles (VLPs) produced by the expression of the prM and E genes were separated from most of the contaminant proteins with 0.2-0.4M NaCl, but still retained their spherical forms and immunogenicity in mice. The column, with a 1 mL bed-volume, concentrated WN-VLPs a minimum of 15 fold from culture supernatants. A heparin analogue, suramin, competitively eluted WN-VLPs, but sulphated polysaccharides, such as heparin, heparin sulfate and dextran sulfate, did not. Furthermore, 2.4 × 10⁹ plaque forming units of WNV and 196 μg of the viral antigens were recovered from 60 mL of infected culture medium at high yields (93% and 96%, respectively). These results indicate that, in addition to conventional methods, Cellufine Sulfate column chromatography is an effective preparation technique for WNV particulate antigens that does not impair the antigen virological characteristics.
authors: Department of Pathology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan. ohtaki@nih.go.jp
authors: Naohiro
authors: N
authors: Ohtaki
authors: Hidehiro
authors: H
authors: Takahashi
authors: Keiko
authors: K
authors: Kaneko
authors: Yasuyuki
authors: Y
authors: Gomi
authors: Toyokazu
authors: T
authors: Ishikawa
authors: Yasushi
authors: Y
authors: Higashi
authors: Masami
authors: M
authors: Todokoro
authors: Takeshi
authors: T
authors: Kurata
authors: Tetsutaro
authors: T
authors: Sata
authors: Asato
authors: A
authors: Kojima
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D051379:Mice; D014771:Virion / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*; D022701:Virosomes / Q000302:isolation & purification; D014902:West Nile virus / Q000302:isolation & purification*
chemical_list: D022701:Virosomes