Refrigerator-temperature stable influenza vaccine compositions

Abstract

Methods and compositions for the optimization and production of refrigerator-temperature stable virus, e.g., influenza, compositions are provided. Formulations and immunogenic compositions comprising refrigerator-temperature stable virus compositions are provided.

What is claimed is: 
     
         1 . A method of making a refrigerator stable influenza virus composition, the method comprising: infection of a population of host eggs or into a population of host cells with influenza viruses; culturing the population of host eggs or population of host cells at an appropriate temperature; recovering influenza viruses in a viral harvest; clarifying the viral harvest by filtration, thereby producing a clarified viral harvest; subjecting the clarified viral harvest to continuous flow centrifugation, thereby producing a further clarified viral harvest; and sterilizing said further clarified viral harvest by filtration. 
     
     
         2 . The method of  claim 1 , wherein the influenza viruses comprise one or more of: an attenuated influenza virus, a cold-adapted influenza virus, a temperature-sensitive influenza virus, or an attenuated cold-adapted temperature-sensitive influenza virus. 
     
     
         3 . The method of  claim 1 , wherein the influenza viruses comprise one or more influenza A virus strains and/or one or more influenza B virus strains. 
     
     
         4 . A refrigerator stable influenza virus composition produced by the method of  claim 1 ,  2 , or  3 . 
     
     
         5 . A refrigerator stable influenza virus composition which comprises at least one strain of influenza virus and one or more members selected from the group consisting of:
 a) 6-8% sucrose   b) 1-2% arginine monohydrochloride;   c) 0.05-0.1% glutamic acid, monosodium monohydrate; and   d) 0.5-2% gelatin hydrolysate.   
     
     
         6 . The refrigerator stable influenza virus composition of  claim 5 , which comprises 6.84% sucrose, 1.21% arginine monohydrochloride, 0.094% glutamic acid, monosodium monohydrate; and 1% gelatin hydrolysate. 
     
     
         7 . The refrigerator stable influenza virus composition of  claim 4 , wherein said composition has a potency loss of less than 1.0 logs when stored at 4° C. for at least 3 months. 
     
     
         8 . The refrigerator stable influenza virus composition of  claim 5 ,  6 , or  7 , wherein said composition has a potency loss of less than 1.0 logs when stored at 4° C. for at least 3 months. 
     
     
         9 . The refrigerator stable influenza virus composition of  claim 8 , wherein said composition further comprises a cold adapted temperature-sensitive attenuated influenza virus. 
     
     
         10 . An immunogenic composition comprising the refrigerator stable influenza virus composition of  claim 7 . 
     
     
         11 . An immunogenic composition comprising the refrigerator stable influenza virus composition of  claim 9 . 
     
     
         12 . A vaccine comprising the immunogenic composition of  claim 10 . 
     
     
         13 . A vaccine comprising the immunogenic composition of  claim 11 . 
     
     
         14 . The method of  claim 1 , wherein the influenza viruses comprise a genetic backbone of one or more of the following influenza viruses: A/Ann Arbor/6/60 and B/Ann Arbor/1/66. 
     
     
         15 . A refrigerator stable influenza virus composition produced by the method of  claim 14 . 
     
     
         16 . An immunogenic composition comprising the refrigerator stable influenza virus composition of  claim 15 . 
     
     
         17 . A vaccine comprising the immunogenic composition of  claim 16 . 
     
     
         18 . A refrigerator stable immunogenic composition comprising at least one live influenza virus, wherein the composition has a potency loss of less than 1.0 logs when stored for a period of 3 months at 4-8° C. 
     
     
         19 . The refrigerator stable immunogenic composition of  claim 18 , wherein the composition has a potency loss of less than 1.0 logs when stored for a period of 12 months at 4-8° C. 
     
     
         20 . The refrigerator stable immunogenic composition of  claim 18  comprising three influenza virus strains. 
     
     
         21 . The refrigerator stable immunogenic composition of  claim 19  comprising three influenza virus strains. 
     
     
         22 . A vaccine comprising the refrigerator stable immunogenic composition of  claim 18 . 
     
     
         23 . A vaccine comprising the refrigerator stable immunogenic composition of  claim 19 . 
     
     
         24 . A vaccine comprising the refrigerator stable immunogenic composition of  claim 20 . 
     
     
         25 . A vaccine comprising the refrigerator stable immunogenic composition of  claim 21 .