Simple and rapid purification of alphaherpesviruses by chromatography on a cation exchange membrane.
A simple and rapid method is described for the purification of two alphaherpesviruses, pseudorabies virus (PrV) and bovine herpesvirus 1, by chromatography on a cation exchange membrane. Cell culture supernatants were passed over a sulfonic-acid modified filter membrane and virions were eluted with a potassium chloride-containing buffer. Over 85% of the virus was eluted within a single fraction and specific infectivity of the resulting virus preparation was over 10-fold higher than that of sucrose gradient-purified virions. Cation exchange was also used for purification of PrV mutants deleted in several glycoproteins which grow in cell culture to titers 10- to 100-fold lower than those obtained by wildtype PrV. For PrV, the presence of non-essential glycoprotein gC, which mediates interaction of virions with cell surface heparin sulfate during attachment, was crucial for the successful purification by cation exchange.
Karger
A
A
Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, Insel Riems, Germany. karger@rie.bfav.de
Bettin
B
B
Granzow
H
H
Mettenleiter
T C
TC
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002845:Chromatography / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D007242:Herpesvirus 1, Bovine / Q000302:isolation & purification*; D011558:Herpesvirus 1, Suid / Q000302:isolation & purification*; D008854:Microscopy, Electron; D013552:Swine