Stable and filterable enveloped virus formulations
Envelope viruses (e.g. Newcastle disease virus (NDV)) are formulated for storage at moderately cold temperatures (e.g. −20° C.). The formulation is an aqueous solution containing the enveloped virus at a concentration of from 10 6 PFU/mL to 10 12 PFU/mL; and a non-reducing saccharide (e.g. sucrose). When the non-reducing saccharide is a disaccharide it is present in the solution at a concentration of from 5% (w/v) to 50% (w/v), and when it is a monosaccharide it is present in the solution at a concentration of from 2.5% (w/v) to 25% (w/v). The solution has an osmotic pressure of about 250 mOs or higher, and has a pH of from 5 to 10.
1. A method for preserving stability of an enveloped virus, comprising storing the virus in aqueous solution for six months or longer at a temperature from −4° C. to −30° C., wherein the aqueous solution consists essentially of:
the enveloped virus at a concentration of from 10 6 PFU/mL to 10 12 PFU/mL;
and a non-reducing saccharide,
wherein the saccharide is a disaccharide and is present in the solution at a concentration of from 5% (w/v) to 50% (w/v) or a monosaccharide and is present in the solution at a concentration of from 2.5% (w/v) to 25% (w/v);
wherein the solution
has an osmotic pressure of about 250 mOs or higher;
has a pH of from 5 to 10;
contains less than 0.1% (w/v) Sodium chloride;
and contains substantially no glutamate or phosphate.
2. The method of claim 1 , wherein the solution further comprises an amino acid selected from lysine or arginine at a concentration from 0.1% (w/v) to 5% (w/v) or lysine and arginine at a combined concentration from 0.1% (w/v) to 5% (w/v).
3. A method for preserving stability of an enveloped virus, comprising storing the virus in aqueous solution for six months or longer at a temperature from −4° C. to −30° C., wherein the aqueous solution comprises:
an enveloped virus at a concentration of from 10 6 PFU/mL to 10 12 PFU/mL;
and a non-reducing saccharide,
wherein the saccharide is a disaccharide and is present in the solution at a concentration of from 5% (w/v) to 50% (w/v) or a monosaccharide and is present in the solution at a concentration of from 2.5% (w/v) to 25% (w/v);
wherein the solution
has an osmotic pressure of about 250 mOs or higher;
has a pH of from 5 to 10;
and contains
substantially no reducing agents antioxidants, glutamate, or phosphate,
and less than:
0.1% (w/v) Sodium chloride;
1% (w/v) Dextran;
0.5% (w/v) Mannitol;
0.1% (w/v) Sorbitol;
0.01% (w/v) Polysorbate;
0.5% (w/v) polyethylene glycol;
0.1 mM Calcium chloride;
0.5% (w/v) Phosphatidyl choline;
0.05% (w/v) Glycine.
4. The method of claim 3 , wherein the solution contains substantially no Sodium chloride, Dextran, Mannitol, Sorbitol, Polysorbate, polyethylene glycol, Calcium chloride, Phosphatidyl choline, and Glycine.
5. The method of claim 3 , wherein the solution further comprises an amino acid selected from lysine or arginine at a concentration from 0.1% (w/v) to 5% (w/v) or lysine and arginine at a combined concentration from 0.1% (w/v) to 5% (w/v).
6. The method of claim 5 , wherein the concentration of lysine and/or arginine is about 1% (w/v).
7. The method of claim 3 , wherein the storage temperature is about −20° C.
8. The method of claim 3 , wherein the virus is a paramyxovirus.
9. The method of claim 8 , wherein the virus is a Newcastle disease virus.
10. The method of claim 3 , wherein the virus concentration is from 1×10 10 PFU/mL to 7×10 10 PFU/mL.
11. The method of claim 3 , wherein the disaccharide concentration is from 7.5% (w/v) to 15% (w/v).
12. The method of claim 11 , wherein the disaccharide concentration is from about 10% (w/v) to about 20% (w/v).
13. The method of claim 3 , wherein the saccharide is sucrose.
14. The method of claim 3 , wherein the saccharide is trehalose.
15. The method of claim 3 , wherein the monosaccharide concentration is from 4% (w/v) to 7% (w/v).
16. The method of claim 15 , wherein the monosaccharide concentration is about 5% (w/v).
17. The method of claim 3 , wherein the osmotic pressure is about 300 mOs.
18. The method of claim 3 , wherein the pH is from 7 to 9.
19. The method of claim 3 , wherein the solution is sterile.
20. The method of claim 3 , wherein the enveloped virus is a Newcastle disease virus and the saccharide is sucrose at a concentration from about 10% (w/v) to about 20% (w/v).
21. The method of claim 5 , wherein the enveloped virus is a Newcastle disease virus, the saccharide is sucrose at a concentration from about 10% (w/v) to about 20% (w/v), and the amino acid is Lysine at a concentration of about 1% (w/v).