VIRUS PURIFICATION AND FORMULATION PROCESS
Disclosed herein is provided a virus purification and formulation process for purifying a flavivirus represented by one of a a Yellow Fever Virus, Japanese Encephalitis virus, Dengue virus, and West Nile virus. The highly purified flavivirus virus product is characterized as having a low level of sucrose without significant virus loss such as that which is typically encountered by prior art virus purification processes. The disclosed process captures and purifies the virus, separating it from the host cell proteins and DNA, and leaving the host cell proteins and DNA behind. The process also can be used to inactivate and/or concentrate the virus sufficiently for use in formulations.
1 . An essentially pure inactivated flavivirus virus preparation which is purified by the process comprising:
a. obtaining a composition suitable for purification from virus-infected cell culture;
b. clarifying the composition obtained in step “a” by removing cells and cell debris from said composition by depth filtration;
c. subjecting the filtrate in step “b” to Benzonase digestion and 0.2 Micron Filtration sufficient to fragment Vera cell DNA followed by filtration;
d. purifying the filtrate obtained in step “c” via CELLUFINE® sulfate chromatography to achieve depyrogenation of said virus in said filtrate;
e. stabilizing and inactivating the viral product obtained in step “d” with an inactivation amount of β-PL; and
f. formulating the product obtained in step bound to alum.