Methods of producing influenza vaccine compositions

Abstract

Methods and compositions for the optimization of production of influenza viruses suitable as influenza vaccines are provided.

1 . (canceled) 
     
     
         2 . A method of making a liquid refrigerator-stable live influenza virus vaccine composition comprising:
 (a) passaging live influenza virus through eggs;   (b) harvesting the live influenza virus of (a), thereby generating a viral harvest;   (c) filtering the viral harvest through a membrane;   (d) adding a stabilizer consisting of arginine and gelatin to the viral harvest; and   (e) replacing some or all of normal allantoic fluid with a buffer, which buffer comprises sucrose, potassium phosphate and glutamate (SPG),   
       thereby producing a liquid refrigerator-stable live influenza virus vaccine composition. 
     
     
         3 . The method of  claim 2 , wherein the eggs are rocked during passage in (a) by a rocking process. 
     
     
         4 . The method of  claim 3 , wherein the rocking process comprises tilting the eggs at a rate of about 1 cycle per minute or less, about 5 cycles per minute or less, or about 10 cycles per minute or less. 
     
     
         5 . The method of  claim 3 , wherein the eggs are rocked for about 12 hours. 
     
     
         6 . The method of  claim 3 , wherein the eggs are rocked for about 24 to 48 hours. 
     
     
         7 . The method of  claim 2 , further comprising a secondary incubation, wherein the eggs are rocked during the secondary incubation. 
     
     
         8 . The method of  claim 3 , wherein a TCID 50  of the rocked eggs is at least 0.4 log greater than a TCID 50  of the same influenza virus passaged through non-rocked eggs. 
     
     
         9 . The method of  claim 2 , wherein at least one influenza virus is selected from: an attenuated influenza virus, a cold-adapted influenza virus, a temperature-sensitive influenza virus, an attenuated cold-adapted influenza virus, a temperature-sensitive cold-adapted influenza virus, an attenuated temperature-sensitive influenza virus, and an attenuated cold-adapted temperature-sensitive influenza virus. 
     
     
         10 . The method of  claim 2 , wherein the influenza virus comprises one or more influenza A viruses and/or one or more influenza B viruses. 
     
     
         11 . The method of  claim 2 , wherein the influenza virus comprises at least two influenza virus strains. 
     
     
         12 . The method of  claim 11 , wherein the influenza virus comprises three influenza virus strains. 
     
     
         13 . The method of  claim 2 , wherein the vaccine composition comprises from 1% (w/v) to 5% (w/v) arginine and from 1% (w/v) to 4% (w/v) gelatin. 
     
     
         14 . The method of  claim 2 , wherein the vaccine composition comprises from 1% to 2% (w/v) arginine, 1% (w/v) gelatin and from 7% (w/v) to 10% (w/v) sucrose. 
     
     
         15 . The method of  claim 2 , wherein the vaccine composition when stored from 2° C. to 8° C. exhibits less than or equal to a 1.0 log potency loss in 6 months or less than or equal to a 0.080 log potency loss per month. 
     
     
         16 . The method of  claim 15 , wherein the potency is measured in fluorescent focus units. 
     
     
         17 . An influenza virus vaccine composition produced by the method of  claim 2 .