Method for the Purification of Alphavirus Replicon Particles

Abstract

Methods of production and purification for viruses and virus-derived vectors, including those related to alphaviruses, are disclosed. In one aspect, methods of purification that subject alphavirus replicon particle preparations to one or more steps of chromatographic purification, such as using an ion exchange resin, are provided. Also disclosed are methods of characterizing alphavirus replicon particles and utilizing these materials for vaccines and gene-based therapeutics

1 . A method for purifying alphavirus replicon particles comprising: a) contacting a preparation containing alphavirus replicon particles with a tentacle ion exchange resin, under conditions and for a time sufficient to bind to said resin; b) removing the portion of the preparation which is not bound to said ion exchange resin from said ion exchange resin; c) eluting the bound alphavirus replicon particles from said ion exchange resin; and d) recovering said replicon particles. 
     
     
         2 . The method according to  claim 1  wherein said tentacle ion exchange resin is a cationic exchange resin. 
     
     
         3 . The method according to  claim 1  wherein said tentacle ion exchange resin is an anionic exchange resin. 
     
     
         4 . A method for purifying alphavirus replicon particles comprising at least two chromatographic purification steps wherein said purification steps are selected from the group consisting of ion exchange chromatography, size exclusion chromatography, hydrophobic interaction chromatography, affinity chromatography. 
     
     
         5 . The method according to  claim 4  wherein said ion exchange chromatography is performed using a tentacle ion exchange resin. 
     
     
         6 . The method according to  claim 4  wherein a first purification step is ion exchange chromatography and a second purification step is size exclusion chromatography.