Composition for the preservation of viruses
View Patent ↗This invention relates to a composition for the preservation of a virus, the composition including a virus, a lipid and a cryoprotectant.
1. A composition for the preservation of a virus, said composition comprising a virus, a polycationic lipid that has a hydrophilic moiety that includes two or more groups derived from positively charged amino acids, and a cryoprotectant,
wherein the polycationic lipid has the following chemical formula:
or a salt thereof,
wherein X is a positively-charged hydrophilic moiety; Y is a spacer having a carbon chain length equivalent to 1 to 30 carbon-carbon single covalent bonds or is absent; and R 1 , R 2 , and R 3 are the same or different acyl groups of a fatty acid.
2. A composition according to claim 1 , wherein the lipid has the following chemical formula:
3. A composition according to claim 1 , wherein the concentration of lipid is in the range from 0.1 μM to 1 mM.
4. A composition according to claim 1 , wherein the concentration of lipid is in the range from 1 μM to 500 μM.
5. A composition according to claim 1 , wherein the concentration of lipid is in the range from 5 μM to 100 μM.
6. A composition according to claim 2 , wherein the concentration of lipid is 10 μM.
7. A composition according to claim 1 , wherein the cryoprotectant is a poly-hydroxy compound.
8. A composition according to claim 7 , wherein the poly-hydroxy compound is a sugar.
9. A composition according to claim 8 , wherein the sugar is sucrose, trehalose, dextrose, lactose, maltose, glucose or any combination of these sugars.
10. A composition according to claim 9 , wherein the sugar is sucrose.
11. A composition according to claim 1 , wherein the concentration of the cryoprotectant is in the range from 0.1 to 20% weight/volume.
12. A composition according to claim 1 , wherein the concentration of the cryoprotectant is in the range from 1 to 10% weight/volume.
13. A composition according to claim 10 , wherein the concentration of sucrose is 8.5% weight/volume.
14. A composition according to claim 1 , wherein the composition further includes a surfactant.
15. A composition according to claim 14 , wherein the surfactant is a non-ionic surfactant.
16. A composition according to claim 15 , wherein the non-ionic surfactant is a molecule that includes an oxyethylene group and a hydroxy group.
17. A composition according to claim 16 , wherein the non-ionic surfactant is polysorbate 80 or polyethylene glycol 400.
18. A composition according to claim 14 , wherein the concentration of the surfactant is in the range from 0.0001% to 10% volume/volume.
19. A composition according to claim 17 , wherein the concentration of polysorbate 80 is in the range from 0.0001% to 1% volume/volume.
20. A composition according to claim 17 , wherein the concentration of polysorbate 80 is 0.005% volume/volume.
21. A composition according to claim 17 , wherein the concentration of polyethylene glycol 400 is in the range from 0.01% to 10% volume/volume.
22. A composition according to claim 17 , wherein the concentration of polyethylene glycol 400 is 0.5% volume/volume.
23. A composition according to claim 1 , wherein the virus is a virus derived from one or more of the group consisting of Adenoviridae, Herpesviridae, Poxviridae, Papovaviridae, Orthohepadnavirus, Parvoviridae, Birnaviridae, Reoviridae, Flaviviridae, Picornaviridae, Togaviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Orthomyxoviridae, and Retroviridae.
24. A composition according to claim 23 , wherein the virus is derived from the Adenoviridae family of viruses.
25. A composition according to claim 24 , wherein the virus is an ovine atadenovirus.
26. A composition according to claim 25 , wherein the virus is OAdV623 or a derivative of OAdV623.
27. A composition according to claim 1 , wherein the concentration of virus in the composition is in the range from 1×10 6 to 1×10 14 virus particles/ml.
28. A composition according to claim 1 , wherein the concentration of virus in the composition is in the range from 1×10 8 to 5×10 12 virus particles/ml.
29. A composition according to claim 1 , wherein the pH of the composition is in the range from 4 to 10.
30. A composition according to claim 1 , wherein the pH of the composition is in the range from 6 to 8.5.
31. A composition according to claim 1 , wherein the virus is storage stable.
32. A composition according to claim 31 , wherein the composition is stored at −200° C. to 0° C.
33. A composition according to claim 31 , wherein the composition is stored at −80° C. to −20° C.
34. A composition according to claim 31 , wherein the composition is stored for 12 months or greater.
35. A composition according to claim 31 , wherein the composition is stored for 3 months or greater.
36. A composition according claim 31 , wherein the composition is stored for 1 week or greater.
37. A composition according to claim 31 , wherein the composition is stored for 1 day or greater.
38. A composition according claim 1 , wherein the virus is stable to freeze-thawing.
39. A method of producing a composition for the preservation of a virus, said method comprising preparing a liquid composition comprising a virus, a polycationic lipid that has a hydrophilic moiety that includes two or more groups derived from positively charged amino acids, and a cryoprotectant,
wherein the polycationic lipid has the following chemical formula:
or a salt thereof,
wherein X is a positively-charged hydrophilic moiety; Y is a spacer having a carbon chain length equivalent to 1 to 30 carbon-carbon single covalent bonds or is absent; and R 1 , R 2 , and R 3 are the same or different acyl groups of a fatty acid.
40. A method according to claim 39 , wherein the lipid has the following chemical formula:
41. A method according to claim 39 , wherein the cryoprotectant is a poly-hydroxy compound.
42. A method according to claim 41 , wherein the poly-hydroxy compound is a sugar.
43. A method according to claim 42 , wherein the sugar is sucrose, trehalose, dextrose, lactose, maltose, glucose or any combination of these sugars.
44. A method according to claim 39 , wherein the composition further includes a surfactant.
45. A method according to claim 44 , wherein the surfactant is a non-ionic surfactant.
46. A method according to claim 45 , wherein the non-ionic surfactant is a molecule that includes an oxyethylene group and a hydroxy group.
47. A method according to claim 46 , wherein the non-ionic surfactant is polysorbate 80 or polyethylene glycol 400.
48. A method according to claim 39 , wherein the virus is a virus derived from one or more of the group consisting of Adenoviridae, Atadenoviridae, Herpesviridae, Poxviridae, Parvoviridae, Papoviridae, Orthohepadnavirus, Parvoviridae, Birnaviridae, Reoviridae, Flaviviridae, Picornaviridae, Togaviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Orthomyxoviridae, and Retroviridae.
49. A method according to claim 48 , wherein the virus is derived from the Adenoviridae family of viruses.
50. A method according to claim 39 , wherein the virus is purified by a method including chromatography or centrifugation.
51. A method according to claim 39 , wherein the composition is formed by combining a solution including the virus and the cryoprotectant with a solution including the lipid.
52. A method according to claim 39 , wherein the virus is storage stable.
53. A method according to claim 39 , wherein the virus is stable to freeze-thawing.
54. A composition for the preservation of a virus, said composition comprising a virus, a polycationic lipid that has a hydrophilic moiety that includes two or more groups derived from positively charged amino acids, and a cryoprotectant,
wherein the virus is storage stable when the composition is stored as a frozen liquid, and
wherein the polycationic lipid has the following chemical formula:
or a salt thereof,
wherein X is a positively-charged hydrophilic moiety; Y is a spacer having a carbon chain length equivalent to 1 to 30 carbon-carbon single covalent bonds or is absent; and R 1 , R 2 , and R 3 are the same or different acyl groups of a fatty acid.
55. A composition for the preservation of a virus, said composition comprising a virus, a polycationic lipid that has a hydrophilic moiety that includes two or more groups derived from positively charged amino acids, and a cryoprotectant,
wherein the virus is stable to freeze-thawing, and
wherein the polycationic lipid has the following chemical formula:
or a salt thereof,
wherein X is a positively-charged hydrophilic moiety; Y is a spacer having a carbon chain length equivalent to 1 to 30 carbon-carbon single covalent bonds or is absent; and R 1 , R 2 , and R 3 are the same or different acyl groups of a fatty acid.