Methods for viral inactivation and other adventitious agents

Abstract

The invention provides for methods of viral inactivation using high temperature short time (HTST) treatment and adjustment of various parameters such that generation of precipitate and depositions of precipitate are reduced and/or minimized.

1 . A method for inactivating virus or adventitious agents in cell culture media while the media maintains suitability for cell culture, said method comprising (a) subjecting the cell culture media to high temperature short time (HTST) treatment; and (b) adjusting one or more parameters selected from the group consisting of pH, calcium level and phosphate level. 
     
     
         2 . The method of  claim 1  further comprising adjusting trace metal concentrations. 
     
     
         3 . The method of  claim 2  wherein the trace metals is selected from the group consisting of iron and copper. 
     
     
         4 . The method of  claim 3  wherein iron and/or copper concentrations are adjusted in the media prior to HTST treatment. 
     
     
         5 . The method of  claim 4  wherein iron and/or copper is removed from the media prior to HTST treatment. 
     
     
         6 . The method of  claim 4  further comprising supplementing iron and/or copper to the media following HTST treatment to a suitable level for cell culture. 
     
     
         7 . The method of  claim 1  wherein the pH is adjusted when the media comprises calcium and phosphate. 
     
     
         8 . The method of  claim 7 , wherein the pH is adjusted in preparing the media prior to HTST treatment to a suitable low level. 
     
     
         9 . The method of  claim 7 , wherein the pH is adjusted by lowering to a suitable level. 
     
     
         10 . The method of  claim 7 , wherein the pH is adjusted to less than about 7.2. 
     
     
         11 . The method of  claim 7 , wherein the pH is adjusted to about 5.0-7.2. 
     
     
         12 . The method of  claim 7 , further comprising adjusting the pH following HTST treatment to a suitable level for cell culture. 
     
     
         13 . The method of  claim 12 , wherein the pH is adjusted to about 6.9-7.2. 
     
     
         14 . The method of  claim 1 , wherein the calcium level is adjusted when the media comprises phosphate. 
     
     
         15 . The method of  claim 14 , wherein the calcium level is reduced. 
     
     
         16 . The method of  claim 15  wherein the calcium level is reduced such that formation of complexes comprised of calcium and phosphate is suppressed. 
     
     
         17 . The method of  claim 15 , wherein calcium is removed from the media prior to HTST treatment. 
     
     
         18 . The method of  claim 15 , wherein the pH is adjusted such that formation of complexes comprised of calcium and phosphate is suppressed. 
     
     
         19 . The method of  claim 14  further comprising adjusting the calcium level following HTST treatment to a suitable level for cell culture. 
     
     
         20 . The method of  claim 1 , wherein the phosphate level is adjusted when the media comprises calcium. 
     
     
         21 - 38 . (canceled)