Purification of infectious bronchitis coronavirus by Sephacryl S-1000 gel chromatography.
A procedure was developed to purify infectious bronchitis virus (IBV) by gel chromatography (GC) with a Sephacryl S-1000 column. Virus samples concentrated by centrifugation were applied to a Sephacryl S-1000 column and eluted by 0.02 M phosphate buffer (pH 7.2) containing 0.15 M NaCl. Virus particles were recovered mainly in the first peak. Purity of the samples was evaluated by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electron microscopy. Using electron microscopy, it was found that there were more spike-rich particles in the virus samples purified by GC than in those purified by sucrose density gradient centrifugation (SDGC). In addition, the hemagglutination unit [log10 (infectivity titer/hemagglutination titer)] of GC-purified virus samples was approximately 10 times lower than that of SDGC-purified virus samples. These results indicate that Sephacryl S-1000 gel chromatography is useful for purification of IBV.
Nagano
H
H
Research Center for Veterinary Science, Kitasato Institute, Chiba, Japan.
Yagyu
K
K
Ohta
S
S
D000818:Animals; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003332:Coronaviridae / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006386:Hemagglutination Tests; D001351:Infectious bronchitis virus / Q000302:isolation & purification* / Q000648:ultrastructure; D008854:Microscopy, Electron