Methods for preparing squalene
An improved method for preparing squalene from a squalene-containing composition, said method comprising the steps of (a) a purification distillation carried out at a temperature T 1 (b) a denaturing distillation carried out at a temperature T 2 ; wherein steps (a) and (b) may be performed in either order; T 1 and T 2 are sufficient to cause squalene to boil; T 2 >T 1 ; and T 2 >200° C.
1. A method for the manufacture of an oil-in-water emulsion comprising an aqueous component, squalene, and 0.01 to 2% by weight polysorbate 80, the method comprising: (i) preparing squalene from a composition comprising squalene from an animal source by a process comprising steps of: (a) a purification distillation carried out at a temperature T 1 ; and (b) a denaturing distillation carried out at a temperature T 2 ; wherein steps (a) and (b) may be performed in either order; T 1 and T 2 are sufficient to cause squalene to boil; T 2 ≥200° C.; T 2 >T 1 ; and T 2 −T 1 is from 10° C. to 300° C.; and
(ii) preparing an emulsion using the squalene prepared in step (i).
2. The method of claim 1 , wherein the squalene prepared in step (i) is kept sterile following distillation treatment and prior to the preparation of the emulsion in step (ii).
3. The method of claim 1 , wherein T 2 −T 1 is from 30° C. to 250° C.
4. A method for the manufacture of a pharmaceutically acceptable composition comprising an oil-in-water emulsion, comprising:
(i) preparing squalene from a composition comprising squalene from an animal source by a process comprising steps of: (a) a purification distillation carried out at a temperature T 1 ; and (b) a denaturing distillation carried out at a temperature T 2 ; wherein steps (a) and (b) may be performed in either order; T 1 and T 2 are sufficient to cause squalene to boil; a T 2 ≥200° C.; T 2 >T 1 ; and T 2 −T 1 is from 10° C. to 300° C.; and
(ii) preparing an emulsion using the squalene prepared in step (i); and
(iii) preparing a pharmaceutically acceptable composition using the emulsion prepared in step (ii).
5. The method of claim 4 , wherein the squalene prepared in step (i) is kept sterile following distillation treatment and prior to the preparation of the emulsion in step (ii).
6. The method of claim 4 , wherein the pharmaceutically acceptable composition is an influenza vaccine and further comprises an influenza virus antigen.
7. The method of claim 6 , wherein the antigen is a hemagglutinin antigen.
8. The method of claim 7 , wherein the hemagglutinin antigen is of a hemagglutinin subtype selected from the group consisting of H2, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15 and H16.
9. The method of claim 6 , wherein the influenza vaccine comprises antigen from one influenza virus strain or more than one influenza virus strain.
10. The method of claim 6 , wherein the influenza virus strain is a reassortant strain.
11. The method of claim 6 , wherein the influenza virus strain is obtained by reverse genetics.
12. The method of claim 6 , wherein the influenza virus strain is grown in egg.
13. The method of claim 6 , wherein the influenza virus strain is grown in cell culture.
14. The method of claim 7 , wherein the cell culture is in a mammalian cell line.
15. The method of claim 14 , wherein the mammalian cell line is MDCK cells.
16. The method of claim 4 , wherein T 2 −T 1 is from 30° C. to 250° C.