SIMPLE AND RAPID PURIFICATION OF ALPHAHERPESVIRUSES BY CHROMATOGRAPHYON A CATION-EXCHANGE MEMBRANE

A simple and rapid method is described for the purification of two alp haherpesviruses, pseudorabies virus (PrV) and bovine herpesvirus 1, by chromatography on a cation exchange membrane. Cell culture supernatan ts were passed over a sulfonic-acid modified filter membrane and virio ns were eluted with a potassium chloride-containing buffer. Over 85% o f the virus was eluted within a single fraction and specific infectivi ty of the resulting virus preparation was over 10-fold higher than tha t 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 obtai ned by wildtype PrV. For PrV, the presence of non-essential glycoprote in gC, which mediates interaction of virions with cell surface heparin sulfate during attachment, was crucial for the successful purificatio n by cation exchange. (C) 1998 Elsevier Science B.V. All rights reserv ed.