Production of poliovirus at high titers for vaccine production
AssigneeCrucell Holland B.V.
InventorJohn Alfred Lewis
Life of this patent
Estimating the term…
Abstract
Provided is a process for the production of poliovirus, comprising the steps of: a) providing a serum-free suspension culture of cells, which are primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences, b) infecting the cells with poliovirus, at a cell density of between 2×106 cells/ml and 150×106 cells/ml, and c) harvesting poliovirus at a time of between 12 and 48 hours after infection.
Claims60
A process for the production of poliovirus at a titer of at least 1010 CCID50/mL from a serum-free suspension cell culture, the process comprising:
- providing a serum-free suspension culture of cells, which cells are primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences;
- infecting the immortalized HER cells in the serum-free suspension culture, at a cell density of between 5×106 cells/mL and 50×106 cells/mL, with poliovirus, at a multiplicity of infection (MOI) of between 0.1 and 3 CCID50/cell;
- propagating the poliovirus in the cells, thus producing poliovirus at a titer of at least 1010 CCID50/mL;
- and
- harvesting a poliovirus preparation from the infected HER cells at a time of between 12 and 48 hours after infection.
A process for producing poliovirus at a titer of at least 1010 CCID50/mL from a serum-free suspension cell culture, the process comprising:
- providing a serum-free suspension culture of cells, wherein the cells are as deposited with the European Collection of Cell Cultures (ECACC), under deposit reference no. 96022940;
- infecting the cells in the serum-free suspension culture, at a cell density of between 5×106 cells/mL and 50×106 cells/mL, with poliovirus at a multiplicity of infection (MOI) of between 0.1 and 3 CCID50/cell;
- propagating the poliovirus in the cells, thus producing poliovirus at a titer of at least 1010 CCID50/mL;
- and
- harvesting a poliovirus preparation from the infected cells at a time of between 12 and 48 hours after infection.
A process for obtaining a poliovirus preparation having a titer of at least 1010 CCID50/mL from a serum-free suspension cell culture, the process comprising:
- providing a serum-free suspension culture of cells, wherein the cells are primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences;
- infecting the immortalized HER cells with poliovirus, at a cell density between 10 ×106 cells/mL and 50×106 cells/mL;
- propagating the poliovirus in the cells at a temperature between 34° C. and 36° C.;
- and
- harvesting the poliovirus preparation at a time between 18 and 30 hours after infection, to obtain the poliovirus preparation having a titer of at least 1010 CCID50/mL.
A process for producing an inactivated polio vaccine, wherein the process comprises:
- providing a serum-free suspension culture of cells, wherein the cells are primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences;
- infecting the immortalized HER cells with poliovirus, at a cell density of between 10×106 cells/mL and 50×106 cells/mL;
- propagating the poliovirus in the cells;
- harvesting a poliovirus preparation having a titer of at least 1010 CCID50/mL at a time of between 12 and 48 hours after infection to obtain a harvested poliovirus preparation;
- purifying the poliovirus from the harvested poliovirus preparation;
- and
- inactivating the purified poliovirus.
A process for producing poliovirus in a serum-free suspension cell culture at a titer of at least 1010 CCID50/mL, the process comprising:
- providing a serum-free suspension culture with primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences;
- infecting the immortalized HER cells in the serum-free suspension culture, at a cell density of between 10×106 cells/mL and 50×106 cells/mL, with poliovirus type 1 at a multiplicity of infection (MOI) of from 0.1 CCID50/cell to 3 CCID50/cell;
- and
- propagating the poliovirus in the infected cells for between 12 and 30 hours after infection at a temperature of between 33° C. and 37° C., thus producing a poliovirus preparation having a titer of at least 1010 CCID50/mL.
A process for producing poliovirus in a serum-free suspension cell culture at a titer of at least 1010 CCID50/mL, the process comprising:
- providing a serum-free suspension culture with primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences;
- infecting the immortalized HER cells in the serum-free suspension culture, at a cell density of about 10×106 cells/mL, with poliovirus type 1 at a multiplicity of infection (MOI) of between one (1) and three (3);
- and
- propagating the poliovirus in the infected cells for between 12 and 48 hours after infection at a temperature of between 34° C. and 36° C., thus producing a poliovirus preparation having a titer of at least 1010 CCID50/mL.
A process for obtaining a poliovirus preparation having a titer of at least 1010 CCID50/mL from a serum-free suspension cell culture, the process comprising:
- providing a serum-free suspension culture of cells, wherein the cells are as deposited with the European Collection of Cell Cultures (ECACC), under deposit reference no. 96022940;
- infecting the cells with poliovirus, at a cell density between 10×106 cells/mL and 50 ×106 cells/mL;
- propagating the poliovirus in the cells;
- and
- harvesting the poliovirus preparation at a time between 18 and 30 hours after infection, to obtain the poliovirus preparation having a titer of at least 1010 CCID50/mL.
A process for producing an inactivated polio vaccine, wherein the process comprises:
- providing a serum-free suspension culture of cells, wherein the cells are as deposited with the European Collection of Cell Cultures (ECACC), under deposit reference no. 96022940;
- infecting the cells with poliovirus, at a cell density of between 10×106 cells/mL and 50 ×106 cells/mL;
- propagating the poliovirus in the cells;
- harvesting a poliovirus preparation having a titer of at least 1010 CCID50/mL at a time of between 12 and 48 hours after infection to obtain a harvested poliovirus preparation;
- purifying the poliovirus from the harvested poliovirus preparation;
- and
- inactivating the purified poliovirus.
A process for producing poliovirus in a serum-free suspension cell culture at a titer of at least 1010 CCID50/mL, the process comprising:
- providing a serum-free suspension culture with cells as deposited with the European Collection of Cell Cultures (ECACC), under deposit reference no. 96022940;
- infecting the cells in the serum-free suspension culture, at a cell density of between 10 ×106 cells/mL and 50×106 cells/mL, with poliovirus type 1 at a multiplicity of infection (MOI) of from 0.1 CCID50/cell to 3 CCID50/cell;
- and
- propagating the poliovirus in the infected cells for between 12 and 30 hours after infection at a temperature of between 33° C. and 37° C., thus producing a poliovirus preparation having a titer of at least 1010 CCID50/mL.
A process for producing poliovirus in a serum-free suspension cell culture at a titer of at least 1010 CCID50/mL, the process comprising:
- providing a serum-free suspension culture with cells as deposited with the European Collection of Cell Cultures (ECACC), under deposit reference no. 96022940;
- infecting the cells in the serum-free suspension culture, at a cell density of about 10×106 cells/mL, with poliovirus type 1 at a multiplicity of infection of between one (1) and three (3);
- and
- propagating the poliovirus in the infected cells for between 12 and 48 hours after infection at a temperature of between 34° C. and 36° C., thus producing a poliovirus preparation having a titer of at least 1010 CCID50/mL.
Dependencies are inferred from the claim text.