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 - 67 . (canceled) 
     
     
         68 . A method of making a liquid refrigerator stable live influenza virus vaccine composition comprising:
 (a) concentrating a live influenza viral harvest by cross flow filtration; and   (b) washing the concentrated viral harvest with a buffer by cross flow filtration,   wherein the composition comprises, from about 5% to about 10% sucrose, about 1% to about 2% arginine, about 1% to about 4% gelatin, and less than 5% normal allentoic fluid, and wherein the composition when stored from about 2° C. to about 8° C. exhibits less than or equal to 1.0 log potency loss in 12 months or less than or equal to 0.080 log potency loss per month.   
     
     
         69 . The method of  claim 68 , wherein the viral harvest is from eggs. 
     
     
         70 . The method of  claim 68 , wherein the viral harvest is from cell culture. 
     
     
         71 . The method of  claim 68 , wherein the influenza viral harvest is concentrated 10× to 200×. 
     
     
         72 . The method of  claim 68 , wherein the concentrated viral harvest is washed with about 5 volumes of buffer, wherein 1 volume equals the volume of the viral harvest prior to concentration. 
     
     
         73 . The method of  claim 68 , wherein the volume after washing is the same as the volume of the concentrated virus before washing. 
     
     
         74 . The method of  claim 68 , wherein the buffer comprises sucrose, potassium phosphate and glutamate (SPG). 
     
     
         75 . The method of  claim 68 , wherein the viral harvest is stabilized prior to concentrating. 
     
     
         76 . The method of  claim 75 , wherein the viral harvest is stabilized by adding a buffer comprising SPG. 
     
     
         77 . The method of  claim 68 , wherein the viral harvest is clarified prior to concentrating. 
     
     
         78 . The method of  claim 68 , wherein the composition comprises from about 1% to about 2% arginine. 
     
     
         79 . The method of  claim 68 , wherein the influenza virus composition comprises about 1% gelatin. 
     
     
         80 . The method of  claim 68 , wherein the composition comprises from about 7% to about 10% sucrose. 
     
     
         81 . The method of  claim 68 , wherein the composition further comprises from about 5 mM to about 200 mM potassium phosphate buffer. 
     
     
         82 . The method of  claim 81 , wherein the potassium phosphate buffer is at pH 7.2. 
     
     
         83 . The method of  claim 81 , wherein the composition comprises about 100 mM potassium phosphate buffer. 
     
     
         84 . The method of  claim 81 , wherein the composition comprises from about 10 mM to about 12 mM potassium phosphate buffer. 
     
     
         85 . The method of  claim 68 , wherein the composition has a pH of 7.2. 
     
     
         86 . The method of  claim 68 , wherein the composition does not comprise EDTA. 
     
     
         87 . The method of  claim 68 , wherein the influenza virus is cold adapted. 
     
     
         88 . The method of  claim 68 , wherein the potency is measured in fluorescent focus units.