Simple and rapid purification of alphaherpesviruses by chromatography on a cation exchange membrane

Karger, Axel GND; Bettin, Barbara GND; Granzow, Harald GND; Mettenleiter, Thomas C. GND

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.

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Karger, Axel / Bettin, Barbara / Granzow, Harald / et al: Simple and rapid purification of alphaherpesviruses by chromatography on a cation exchange membrane. 1998.

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