Compositions, methods and uses for inducing viral growth
Embodiments herein report methods, compositions and uses for inducing and/or accelerating viral growth. In certain embodiments, methods, compositions and uses generally relate to copolymer compositions for inducing viral growth, reducing lag time and/or increasing viral plaque size. In other embodiments, methods, compositions and uses of copolymer compositions can be for inducing flaviviral growth, reducing lag in growth and/or increasing plaque size.
1. A method for culturing viruses for production and manufacture, the method comprising:
growing host cells;
introducing to the host cells a composition comprising media for growing viral cultures comprising one or more ethylene oxide propylene oxide (EO-PO) block copolymers before, during, or after viral infection of the host cells; the one or more EO-PO block copolymers comprising poloxamer 407, poloxamer 403, or a combination thereof, wherein the concentration of the EO-PO block copolymer is from 0.001% to 3.0% (w/v);
introducing viruses to the host cells before, during or after introduction of the composition;
incubating the host cells, the viruses, and the media for a predetermined period to culture the viruses;
separating the media from the host cells; and
harvesting the cultured viruses from the media.
2. The method of claim 1 , wherein separating the media from the host cells further comprises removing part of the media from the culture and introducing fresh media to the culture for further viral growth.
3. The method of claim 1 , wherein growth of the viruses is increased compared to viral population growth in cultures without the one or more EO-PO block copolymers.
4. The method of claim 1 , wherein plaque size is increased compared to viral plaque size of cultures without the one or more EO-PO block copolymers.
5. The method of claim 1 , further comprising removing growth media containing the viruses after introduction of the virus to the host cells and collecting the viruses.
6. The method of claim 1 , wherein at least one of the one or more EO-PO block copolymers comprises poloxamer 407.
7. The method of claim 1 , wherein the media for growth comprises Dulbecco's Modified Eagle Medium (DMEM).
8. The method of claim 1 , wherein the viruses comprise live, attenuated viruses for use in vaccines.
9. The method of claim 1 , wherein the viruses are selected from the group consisting of Flavivirus, Togavirus, Coronavirus, Filovirus, Paramyxovirus, Orthomyxovirus, Bunyavirus, Arenavirus, Retrovirus, Hepadnavirus, Pestivirus, Herpes virus, and Poxvirus.
10. The method of claim 1 , wherein the viruses are flavivirus viruses.
11. The method of claim 1 , wherein the viruses are Poxvirus viruses.
12. The method of claim 1 , wherein when harvested, the viral cultures have a titer between about 1×10 6 pfu/ml to about 1×10 9 pfu/ml.
13. The method of claim 1 , wherein the host cells comprise host cells grown to confluency at a density of between about 1×10 4 and about 1×10 5 cells per cm 2 .
14. The method of claim 1 , wherein the host cells comprise Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
15. The method of claim 1 , wherein the EO-PO block copolymer comprises poloxamer 407, and the concentration of poloxamer 407 is from 0.063% to 3.0% (w/v).
16. A method for culturing flaviviruses for production and manufacture, the method comprising:
growing host cells to near confluency;
introducing to the host cells a composition comprising media for growing viral cultures and one or more ethylene oxide propylene oxide (EO-PO) block copolymers before, during, or after viral infection of the host cells; the one or more EO-PO block copolymers comprising poloxamer 407, poloxamer 403, or a combination thereof, wherein the concentration of the EO-PO block copolymer is from 0.001% to 3.0% (w/v);
introducing the flaviviruses to the host cells before, during or after introduction of the composition;
incubating the host cells, the flaviviruses, and the growth media for about 1 hour to about 5 hours to culture the flaviviruses;
separating the media from the host cells; and
harvesting the cultured flaviviruses from the media.
17. The method of claim 16 , removing growth media containing the flaviviruses after introduction of the virus to the host cells and collecting the flaviviruses.
18. The method of claim 16 , wherein the host cells comprise Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
19. The method of claim 16 , wherein the flaviviruses comprise dengue virus, West Nile virus, yellow fever virus, Japanese encephalitis virus, St. Louis encephalitis virus, or tick-borne encephalitis virus.
20. The method of claim 16 , wherein the flaviviruses comprise live, attenuated flaviviruses.
21. The method of claim 16 , wherein the flaviviruses comprise dengue viruses.
22. The method of claim 20 , wherein the live, attenuated flaviviruses are flavivirus chimeras.
23. The method of claim 16 , wherein the growth media comprises Dulbecco's Modified Eagle Medium (DMEM).
24. A method for culturing dengue viruses for production and manufacture, the method comprising:
growing host cells to near confluency;
introducing to the host cells a composition comprising media for growing dengue viral cultures and one or more ethylene oxide propylene oxide (EO-PO) block copolymers before, during, or after viral infection of the host cells; the one or more EO-PO block copolymers comprising poloxamer 407, poloxamer 403, or a combination thereof, wherein the concentration of the EO-PO block copolymer is from 0.001% to 3.0% (w/v);
introducing the dengue viruses to the host cells before, during or after introduction of the composition;
incubating the host cells, the dengue viruses, and the growth media to culture the dengue viruses;
separating the media from the host cells; and
harvesting the cultured dengue viruses from the media.
25. The method of claim 24 , wherein incubating comprises incubating for about 1hour to about 5 hours.
26. The method of claim 24 , wherein separating the media from the host cells further comprises removing part of the media from the culture and introducing fresh media to the culture for further dengue viral growth.
27. The method of claim 24 , further comprising increasing the growth of the dengue viruses compared to dengue viral population growth in cultures without the one or more EO-PO block copolymers.
28. The method of claim 24 , further comprising increasing plaque size compared to dengue viral plaque size of cultures without the one or more EO-PO block copolymers.
29. The method of claim 24 , removing growth media containing the viruses after introduction of the virus to the host cells and collecting the viruses.
30. The method of claim 24 , wherein the dengue viruses are live, attenuated dengue viruses.
31. The method of claim 24 , wherein the dengue viruses are dengue-dengue chimeras.
32. The method of claim 24 , wherein the host cells comprise Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
33. The method of claim 24 , wherein the growth media comprises Dulbecco's Modified Eagle Medium (DMEM).
34. A composition for growing flaviviruses comprising:
a flavivirus culture;
one or more ethylene oxide propylene oxide (EO-PO) block copolymers, the EO-PO block copolymers comprising poloxamer 407, poloxamer 403, or a combination thereof, wherein the concentration of the EO-PO block copolymer is from 0.001% to 3.0% (w/v);
a host cell culture; and
growth media, wherein the EO-PO block copolymers increase growth of the flaviviruses in the host cell culture.
35. The composition of claim 34 , wherein the flavivirus cultures comprises live, attenuated flaviviruses.
36. The composition of claim 34 , wherein the flavivirus cultures comprise dengue viruses, West Nile viruses, yellow fever viruses, Japanese encephalitis viruses, St. Louis encephalitis viruses, or tick-borne encephalitis viruses.
37. The composition of claim 34 , wherein the flavivirus cultures comprise chimeric flaviviruses.
38. The composition of claim 34 , wherein the host cell culture comprises Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
39. The composition of claim 34 , wherein the growth media comprises Dulbecco's Modified Eagle Medium (DMEM).
40. A composition for growing dengue viruses comprising:
a dengue virus culture;
one or more ethylene oxide propylene oxide (EO-PO) block copolymers, the EO-PO block copolymers comprising poloxamer 407, poloxamer 403, or a combination thereof, wherein the concentration of the EO-PO block copolymer is from 0.001% to 3.0% (w/v);
a host cell culture; and
growth media;
wherein the EO-PO block copolymers accelerate dengue virus growth in the host cell culture.
41. The composition of claim 40 , wherein the dengue viruses comprises live, attenuated dengue viruses.
42. The composition of claim 40 , wherein the dengue viruses comprise chimeric dengue viruses.
43. The composition of claim 40 , wherein the host cell culture comprises Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
44. The composition of claim 40 , wherein the growth media comprises Dulbecco's Modified Eagle Medium (DMEM).
45. The composition of claim 40 , wherein EO-PO block copolymer comprises poloxamer 407, and the concentration of poloxamer 407 is from 0.063% to 3.0% (w/v).