Purification and concentration of non-infectious West Nile virus-like particles and infectious virions using a pseudo-affinity Cellufine Sulfate column.

摘要

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 0.2-0.4 M 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, 2.4x109 plaque forming units of WNV and 196 micro g 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.