Methods for the production of hemagglutinating virus of Japan and adenovirus using cultured cells
Inventors
Yano, Takahiro • Nagasawa, Tetsuji • Temma, Akiko • Suzuki, Naho • Miyaji, Kazue
Assignees
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Publication Number
US-8012749-B2
Publication Date
2011-09-06
Expiration Date
Abstract
A novel cell suited for mass production of Hemagglutinating Virus of Japan (HVJ), a method for obtaining the cell and use of the cell are disclosed. The human cell is originated from a transformed human kidney cell line, the doubling time thereof in logarithmic growth phase in suspension culture in a serum-free medium is not more than 40 hours, the cell has a freeze-recovery property, the maximum density of viable cells in suspension culture is not less than 106 cells/mL, and HVJ can grow in the cell. The method for obtaining the human cell comprises the steps of suspension-culturing a human transformed kidney cell line in a serum-free medium, and cloning the grown cells; and selecting, from the cloned cells, a cell whose doubling time in logarithmic growth phase in suspension culture in a serum-free medium is not more than 40 hours, which has a freeze-recovery property, whose maximum density of viable cells in suspension culture is not less than 106 cells/mL, in which HVJ can grow.
Core Innovation
The invention provides an isolated human transformed kidney cell line, identified as GIC20 and deposited under accession number FERM BP-10399. The cell line is preferably a 293-derived, suspension-growth selected human transformed kidney cell line capable of supporting HVJ propagation.
GIC20 is characterized for suspension growth in serum-free medium, including a doubling time of 40 h or less in log phase, maximum viable cell density of 10^6 cells/mL or more, and freeze-recovery property. These culture performance features are described as relevant to HVJ production and as enabling storage and transport of the cell line.
The disclosed system is applied to mass production of HVJ virus using GIC20 cells, including characterization using HVJ focus-forming units (FFU) and assays such as neuraminidase (NA) activity and hemagglutination (HA) tests. The disclosure also states an ability to produce other viruses, including adenovirus, using the same GIC20 cells, and indicates capability for adenovirus vector production.
Claims Coverage
The document presents three independent claims: one to the specific deposited cell line GIC20 and two to methods for producing HVJ virus and adenovirus. The claim set focuses on the deposited identity of the cell line and its use as a production platform by infecting, growing in culture medium, and recovering the virus.
Deposited GIC20 cell line accession
A cell identified as GIC20, deposited under accession number FERM BP-10399.
HVJ production using infected GIC20 cells
A method of producing Hemagglutinating Virus of Japan (HVJ virus) comprising infecting GIC20 cells, deposited under accession number FERM BP-10399, with said HVJ virus; growing said HVJ virus in said GIC20 cells in culture medium; and recovering said HVJ virus.
Adenovirus production using infected GIC20 cells
A method of producing adenovirus comprising infecting GIC20 cells, deposited under accession number FERM BP-10399, with said adenovirus; growing said adenovirus in said GIC20 cells in culture medium; and recovering said adenovirus.
Overall, the claim coverage centers on a specific deposited transformed human kidney cell line (GIC20, FERM BP-10399) and its use for HVJ virus and adenovirus production through infecting, growing in culture medium, and recovering the respective virus.
Stated Advantages
Serum-free medium reduces serum-derived contaminants.
Suspension high-density culture supports bioreactor scale-up.
Freeze-recovery aids storage and transport.
Documented Applications
Mass production of Hemagglutinating Virus of Japan (HVJ virus) using GIC20 cells in culture, with HVJ FFU and additional assays such as neuraminidase (NA) activity and hemagglutination (HA) tests.
Production of other viruses, including adenovirus, and stated capability for adenovirus vector production using GIC20 cells.
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