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 related 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 manufacturing viruses, 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 one or more EO-PO block copolymers is from 0.001% to 3.0%;
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;
separating the media from the host cells; and
harvesting the 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 expansion.
3 . The method of claim 1 , wherein the growth media containing the viruses is removed daily for between one week to about three weeks after introduction of the virus to the host cells.
4 . The method of claim 1 , wherein at least one of the one or more EO-PO block copolymers comprises poloxamer 407.
5 . The method of claim 1 , wherein the media for growth comprises Dulbecco's Modified Eagle Medium (DMEM).
6 . The method of claim 1 , wherein the viruses comprise live, attenuated viruses for use in vaccines.
7 . 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.
8 . 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.
9 . The method of claim 1 , wherein the concentration of the one or more EO-PO block copolymers is from 0.063% to 3.0%.
10 . A method for manufacturing flaviviruses, the method comprising:
growing adherent host cells to near confluency;
introducing to the adherent 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 one or more EO-PO block copolymers is from 0.001% to 3.0%;
introducing the flaviviruses to the adherent host cells before, during or after introduction of the composition;
incubating the adherent host cells, the flaviviruses, and the growth media for about 1 hour and about 5 hours;
separating the media from the adherent host cells; and
harvesting the flaviviruses from the media.
11 . The method of claim 10 , wherein the growth media containing the flaviviruses is removed daily for between one week to about three weeks after introduction of the virus to the host cells.
12 . The method of claim 10 , wherein the adherent host cells comprise Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
13 . The method of claim 10 , wherein the flaviviruses comprise dengue virus, West Nile virus, yellow fever virus, Japanese encephalitis virus, St. Louis encephalitis virus, or tick-borne encephalitis virus.
14 . A composition for growing flaviviruses comprising:
a flavivirus culture;
one or more ethylene oxide propylene oxide CEO-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 0.063% to 3.0%;
an adherent host cell culture; and
growth media, wherein the EO-PO block copolymers increase expansion of the flaviviruses in the host cell culture.
15 . The composition of claim 14 , wherein flavivirus cultures comprises live, attenuated flaviviruses.
16 . The composition of claim 14 , 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.
17 . The composition of claim 14 , wherein the flavivirus cultures comprise chimeric flaviviruses.
18 . The composition of claim 14 , wherein the adherent host cell culture comprises Vero cells (African green monkey Vero cells), LLC-MK2 cells, or C6/36 mosquito cells.
19 . The composition of claim 14 , wherein the growth media comprises Dulbecco's Modified Eagle Medium (DMEM).