Methods of Producing Influenza Vaccine Compositions
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.