Methods for purifying viruses

Abstract

The invention provides methods for purifying a virus from impurities in an aqueous medium.

What is claimed is:  
     
         1 . A method for purification of a virus preparation comprising: 
 a) subjecting the virus preparation to anion-exchange chromatography, wherein the virus is eluted from an anion-exchange chromatographic medium; and    b) subjecting the virus product of step A to size exclusion chromatography, wherein the virus is eluted from a size exclusion chromatographic medium.    
     
     
         2 . The method of    claim 1   , wherein the virus preparation is a cell lysate.  
     
     
         3 . The method of    claim 2   , wherein the cell lysate is filtered before step a.  
     
     
         4 . The method of    claim 1   , wherein the virus is a recombinant adenovirus.  
     
     
         5 . The method of    claim 1   , wherein the anion-exchange medium is FRACTOGEL™-DEAE.  
     
     
         6 . The method of    claim 1   , wherein the size exclusion medium is Superdex-200.  
     
     
         7 . The method of    claim 1   , wherein the size-exclusion medium is provided in a column prepared as a salt gradient decreasing in ionic strength from the top of the column towards the bottom, the top of the column having a buffer having an ionic strength substantially identical to that of the product of step a.  
     
     
         8 . The method of    claim 1    wherein the anion exchange medium comprises diethylaminoethyl groups on a cross-linked agarose, cellulose, polyacrylamide or polystyrene backbone.  
     
     
         9 . The method of    claim 1   , wherein the size-exclusion medium comprises a cross-linked polysaccharide.  
     
     
         10 . The method of    claim 9   , wherein the cross-linked polysaccharide is a composite of cross-linked agarose and dextran.  
     
     
         17 . The method of    claim 1    wherein the virus is ACN53.  
     
     
         18 . The method of    claim 1   , wherein the anion exchange chromatographic medium is extensively washed before application of the virus preparation.  
     
     
         19 . A virus purified by the method of    claim 1   .