Production of adenoviral vectors using serum-free suspension cell culture in a hollow fiber system

Abstract

The present invention relates to methods for the production of high titers of serum-free lytic viruses in a hollow fiber cartridge capillary system. The invention further relates to methods of infecting target cells at high multiplicity and for producing high concentrations of transduced target cells.

What is claimed is:  
     
         1 . A method for the production of lytic virus, comprising 
 (a) inoculating host cells into a serum-free liquid medium in a capillary system,    (b) infecting the host cells with lytic virus,    (c) culturing the infected host cells, and    (d) harvesting the lytic virus from the culture medium.    
     
     
         2 . The method of  claim 1 , wherein the virus is replication defective.  
     
     
         3 . The method of  claim 1 , wherein the virus is replication competent.  
     
     
         4 . The method of  claim 1 , wherein the virus is an adenovirus.  
     
     
         5 . The method of  claim 1 , wherein the virus is a parvovirus.  
     
     
         6 . The method of  claim 5 , wherein the virus is an adeno-associated virus.  
     
     
         7 . The method of  claim 1 , wherein the virus is herpes simplex virus, polio virus, or papillomavirus.  
     
     
         8 . The method of  claim 1  wherein the host cells are 293 cells.  
     
     
         9 . The method of  claim 1 , wherein the host cells are CCL81.1, Vero, HEL or BHK cells.  
     
     
         10 . The method of  claim 1 , wherein the capillary system is a hollow fiber cartridge system.  
     
     
         11 . The method of  claim 1 , wherein the virus is a viral vector.  
     
     
         12 . The method of  claim 1 , wherein the viral vector encodes a therapeutically effective product.  
     
     
         13 . The method of  claim 1 , wherein the host cells have been adapted to grow in a serum-free medium.  
     
     
         14 . The method of  claim 1 , wherein the host cells were stored in under serum-free conditions.  
     
     
         15 . The method of  claim 14 , wherein the host cells were stored in 10% DMSO.  
     
     
         16 . The method of  claim 1 , wherein viral infection is carried out at a cellular lactate production rate of 500 mg/day.  
     
     
         17 . The method of  claim 1 , wherein the virus is harvested at a cellular lactate production rate of between 1000 and 1500 mg/day.  
     
     
         18 . The method of  claim 1 , wherein the virus used to inoculate the host cells are substantially isolated from cellular material.  
     
     
         19 . The method of  claim 1 , wherein the virus is harvested from the host cells.  
     
     
         20 . The method of  claim 1 , wherein the virus is harvested from the extracapillary space of the capillary system.