Preparation of influenza virus vaccine antigens
A number of improvements for preparing vaccine antigens from disintegrated influenza viruses are disclosed. A splitting step can be followed by detergent exchange. Splitting can take place in the presence of a buffer with a higher ionic strength and/or in the presence of phosphate buffer.
1. A method for disrupting influenza virions, the method comprising steps of:
(i) obtaining a composition comprising influenza virions prepared from a cell culture,
(ii) inactivating the influenza virions to obtain inactivated virions, and
(iii) splitting the inactivated virions in the presence of a buffer having an ionic strength of from 200 mM to 800 mM, wherein the buffer comprises phosphate.
2. The method of claim 1 , wherein the influenza virions are from an influenza A virus.
3. The method of claim 1 , wherein the influenza virions are from an influenza A virus of H5 hemagglutinin subtype.
4. The method of claim 1 , wherein the cell culture is a mammalian cell culture or an avian cell culture.
5. A method for manufacturing an influenza vaccine, the method comprising steps of:
carrying out the method of claim 1 ,
obtaining a bulk antigen preparation from the disrupted virions of step (iii), and
formulating the bulk antigen preparation into an influenza vaccine.
6. The method of claim 5 , wherein the influenza virions are from an influenza A virus.
7. The method of claim 5 , wherein the influenza virions are from an influenza A virus of H5 hemagglutinin subtype.
8. The method of claim 5 , wherein the cell culture is a mammalian cell culture or an avian cell culture.
9. The method of claim 5 , wherein the influenza vaccine contains less than 10 ng host cell DNA.
10. The method of claim 5 , wherein the influenza vaccine is a monovalent or trivalent influenza vaccine.
11. The method of claim 5 , wherein the influenza vaccine comprises hemagglutinin from at least one B strain.
12. The method of claim 5 , wherein the influenza vaccine comprises from about 3.75 μg to about 15 μg hemagglutinin, per strain, per dose.
13. The method of claim 5 , wherein the step of formulating comprises combining the bulk antigen preparation with an adjuvant.
14. The method of claim 13 , wherein the adjuvant is an oil-in-water emulsion.
15. The method of claim 1 , wherein the buffer has an ionic strength of at least 300 mM.
16. The method of claim 1 , wherein the buffer has an ionic strength of at least 400 mM.
17. The method of claim 1 , wherein the buffer has an ionic strength of at least 500 mM.
18. The method of claim 1 , wherein the buffer has a pH of from 6.5 to 8.5.
19. The method of claim 1 , wherein the buffer has a pH of from 7.0 to 8.0.
20. The method of claim 1 , wherein the buffer has a pH of from 7.0 to 7.8.
21. The method of claim 1 , wherein the method results in a higher than 20-fold increase in antigen yield, when compared to a method for disrupting influenza virions comprising corresponding steps (i)-(iii), except that step (iii) is performed in the presence of a Tris buffer.
22. The method of claim 1 , wherein the method results in an increased antigen yield, when compared to a method for disrupting influenza virions comprising corresponding steps (i)-(iii), except that the buffer in step (iii) has an ionic strength of less than 200 mM.