Enveloped virus vaccine and method for production
View Patent ↗The present invention provides methods of production of a purified enveloped virus antigen. In particular, it provides purified Ross River Virus (RRV) antigens, and vaccines comprising purified, inactivated Ross River Virus (RRV) antigen.
1. A method for production of purified Ross River Virus antigen, comprising the steps of infecting a cell culture with Ross River Virus; incubating said cell culture to propagate said virus; harvesting the virus produced; filtering the harvested virus with a first filter having a pore size of between 0.3 and 1.5 μm; filtering the harvested virus with a second filter having a pore size of between 0.1 and 0.5 μm; and purifying the virus antigen.
2. The method according to claim 1 , wherein the cell culture comprises VERO cells that have been grown in serum free medium.
3. The method according to claim 1 , wherein said second filtering step is PERFORMED on a filter having a pore size of about 0.2 μm.
4. The method according to claim 1 , wherein the first and second filtering steps reduce cellular protein and nucleic acid contaminants.
5. A method far the production of a purified Ross River Virus preparation, comprising the steps of infecting a cell culture with Ross River Virus; incubating said cell culture to propagate said virus; harvesting the virus produced; filtering the harvested virus with a first filter having a pore size of between about 0.3 and about 1.5 μm; filtering the harvested virus with a. Second filter having a pore size of between 0.1 and 0.5 μm; treating the filtered virus with a nucleic acid degrading agent; and purifying the virus.
6. The method according to claim 5 , wherein the cell culture comprises VERO cells that have been grown in a serum free medium.
7. The method according to claim 5 , wherein said second filtering step is performed on a filter having a pore size of about 0.2 μm.
8. The method according to claim 5 , wherein the nucleic acid degrading agent is an enzyme having DNase and Kinase activity.
9. The method according to claim 5 , wherein said filtered virus is further treated with a virus inactivating agent.
10. The method according to claim 5 , wherein said preparation is substantially free of contaminating proteins from said cell culture and has less than about 10 pg cellular nucleic acid/μg virus antigen.
11. A method for production of an immunogenic composition comprising purified, inactivated Ross River Virus antigen, comprising the steps of infecting a cell culture with Ross River Virus; incubating said cell culture to propagate said virus; harvesting the virus produced; filtering the harvested virus with a first filter having a pore size of between about 0.3 and about 1.5 μm; filtering the harvested virus with a second filter having a pore size of between 0.1 and 0.5 μm; treating the filtered virus with a nucleic acid degrading agent and a virus inactivating agent; purifying the virus; and formulating the purified virus in an immunogenic composition.
12. The method of claim 11 , wherein said first filter is based on a positively charged matrix and said second filter is based on a hydrophilic matrix.
13. The method of claim 1 , further comprising treating the filtered virus with a nucleic acid degrading agent.
14. The method of claim 1 , wherein said first filter is based on a positively charged matrix and said second filter is based on a hydrophilic matrix.
15. The method of claim 5 , wherein said first filter is based on a positively charged matrix and said second filter is based on a hydrophilic matrix.
16. A method according to any of claims 1 , 5 , or 11 wherein the method is used for large scale production.