Bluetongue virus vaccine and immunogenic compositions, methods of use and methods of producing same
View Patent ↗Provided are immunogenic and vaccine compositions and methods for their preparation and use, which compositions are effective in protecting against, minimizing the severity of, preventing, and/or ameliorating infection of ruminants with Bluetongue virus. Administration to an animal of at least one dose of an adjuvanted and twice inactivated Bluetongue virus composition as disclosed herein is effective in providing immunity to the animal and protection from infection with Bluetongue virus, thereby reducing the severity of and/or preventing disease caused by one or more strains or serotypes of Bluetongue virus.
1. A method of producing inactivated whole Bluetongue virus (BTV) that protects animals from viremia, the method comprising the steps of:
a) treating the BTV with an inactivating agent using a 1:10 ratio of inactivating agent to BTV, while the BTV remains in a complete culture;
b) homogenizing the inactivating agent/BTV mixture of step a) for at least 15 minutes;
c) decanting the mixture of step b) into a container and agitating the mixture for about 24 hours;
d) treating the agitated mixture resultant from step c) with an inactivating agent using a 1:20 ratio of inactivating agent to BTV therein;
e) homogenizing the inactivating agent/BTV mixture of step d) for at least 15 minutes;
f) decanting the mixture of step e) into a container and agitating the mixture for about 48 hours; and
g) neutralizing the inactivating agent, and adjusting the final pH to about 7.2, if needed;
wherein the method results in inactivation of the BTV while maintaining the immunogenicity of the BTV.
2. The method of claim 1 , wherein the inactivating agent is binary ethyleneimine (BEI).
3. The method of claim 1 , wherein the final concentration of inactivating agent in step a) is about 10 mM.
4. The method of claim 1 , wherein the final concentration of inactivating agent in step d) is about 5 mM.
5. The method of claim 1 , wherein the whole Bluetongue virus is serotype 4.