Removal of contaminating viruses from aav preparations

Abstract

The present invention relates to a separation of viruses of an essentially spherical shape from viruses with a rod-like shape that are comprised in a sample, wherein the sample comprising the viruses is subjected to filtration.

1 . A method for separating a population of parvoviral virions from a population of baculoviral virions, comprising filtrating a sample comprising populations of parvoviral and baculoviral virions over a filter in tangential flow filtration modus, wherein the filter has a nominal pore size of 30-40 nm. 
     
     
         2 . The method according to  claim 1 , wherein the sample, prior to filtration, is prepurified using a method selected from the group consisting of a density gradient, a prefiltration, a chromatography step, and combinations of these methods, resulting in a pre-purified sample. 
     
     
         3 . The method according to  claim 1 , wherein the parvovirus is an adeno-associated virus. 
     
     
         4 . The method according to  claim 1 , wherein the baculovirus is Autographa californica multicapsid nucleopolyhedrovirus (AcmNPV). 
     
     
         5 . The method according to  claim 1 , wherein the filter is a virus filter or an ultrafilter. 
     
     
         6 . The method according to  claim 1 , wherein the filter is a surface and/or a depth filter. 
     
     
         7 . The method according to  claim 1 , wherein the filter comprises a material selected from the group consisting of cuprammonium-regenerated cellulose, polyvinylidenefluoride, polysulfone, polytetrafluoroethylene, polypropylene, modified or unmodified polyethersulfone, cellulose acetate, cellulose nitrate, polyamide and regenerated cellulose. 
     
     
         8 . The method according to  claim 1 , wherein the filter has a nominal pore size of 32-38 nm. 
     
     
         9 . The method according to  claim 1 , wherein the filter comprises cuprammonium-regenerated cellulose. 
     
     
         10 . The method according to  claim 9 , wherein the filter comprises cuprammonium-regenerated cellulose hollow fibers. 
     
     
         11 . The method according to  claim 10 , wherein the filter is a Planova 35 membrane. 
     
     
         12 . The method according to  claim 2 , wherein the prepurified sample obtained in  claim 2  is subjected to filtration using two or more filters. 
     
     
         13 . The method according to  claim 2 , wherein the chromatography affinity chromatography and/or ion-exchange chromatography. 
     
     
         14 . The method according to  claim 2 , wherein the prefiltration is effected by filtering with a pore size of 70-200 nm. 
     
     
         15 . The method according to  claim 1 , wherein the pH of the sample is 6 to 10. 
     
     
         16 . The method according to  claim 15 , wherein the pH of the sample is 7 to 9. 
     
     
         17 . The method according to  claim 15 , wherein the pH of the sample is 7.5 to 8.5. 
     
     
         18 . The method according to  claim 1 , wherein 1 to 200 ml of sample is filtered per 1 cm 2  of virus filter surface. 
     
     
         19 . The method according to  claim 18 , wherein 80 to 120 ml of sample is filtered per 1 cm 2  of virus filter surface.