Reverse genetics methods for virus rescue

Abstract

A method for rescuing a virus by reverse genetics is provided in which cells are added after transfection.

1 - 28 . (canceled) 
     
     
         29 . A method for preparing a viral composition, the method comprising the steps of:
 (i) transfecting a culture eukaryotic cells with at least one expression construct encoding a viral RNA molecule;   (ii) adding untransfected eukaryotic cells to the transfected cells of step (i) to provide a mixture of cells, wherein the untransfected cells are of the same type as the cells of step (i);   (iii) culturing the mixture of cells from step (ii) so as to produce viruses in the culture; and,   (iv) harvesting viruses, proteins or combination thereof from the culture obtained in step (iii), so as to obtain a viral composition.   
     
     
         30 . The method of  claim 29 , further comprising a step of:
 (v) purifying the viruses from step (iv) so as to obtain purified virions.   
     
     
         31 . The method of  claim 30 , wherein the virions obtained in step (v) is whole virions. 
     
     
         32 . The method of  claim 30 , further comprising inactivating the viruses or the purified virions. 
     
     
         33 . The method of  claim 31 , further comprising packaging the whole virions in a sterile container. 
     
     
         34 . The method of  claim 33 , wherein the viral composition further comprises an adjuvant. 
     
     
         35 . A kit comprising the whole virions of  claim 33 . 
     
     
         36 . The kit of  claim 34 , further comprising an adjuvant component provided in a separate container. 
     
     
         37 . The method of  claim 30 , further comprising a step of:
 treating the purified virions with a detergent so as to obtain split virions.   
     
     
         38 . The method of  claim 37 , further comprising inactivating the split virions. 
     
     
         39 . The method of  claim 37 , further comprising packaging the split virions in a sterile container. 
     
     
         40 . The method of  claim 39 , wherein the viral composition further comprises an adjuvant. 
     
     
         41 . A kit comprising the split virions of  claim 37 . 
     
     
         42 . The kit of  claim 40 , further comprising an adjuvant component provided in a separate container. 
     
     
         43 . The method of  claim 29 , wherein the proteins of step (iv) are one or more surface antigens. 
     
     
         44 . The method of  claim 43 , further comprising a step of:
 purifying the one or more surface antigens so as to obtain purified surface antigens.   
     
     
         45 . The method of  claim 43 , further comprising inactivating the surface antigens. 
     
     
         46 . The method of  claim 44 , further comprising packaging the purified surface antigens in a sterile container. 
     
     
         47 . The method of  claim 44 , wherein the viral composition further comprises an adjuvant. 
     
     
         48 . A kit comprising the surface antigens of  claim 46 . 
     
     
         49 . The kit of  claim 47 , further comprising an adjuvant component provided in a separate container.