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US 6,146,873 AGranted
Production of orthomyxoviruses in monkey kidney cells using protein-free media
AssigneeBaxter Aktiengesellschaft
InventorsOtfried Kistner, Noel Barrett, Wofgang Mundt, Friedrich Dorner
Life of this patent
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Abstract
Viruses from the family Orthomyxoviridae, particularly influenza virus, can grown in monkey kidney cells, particularly Vero Cells, after passaging the cells in a serum-free or protein-free medium. The use of a proteolytic enzyme, especially trypsin, also aids in the propagation of the virus. The method allows for the virus to be produced to be used in a vaccine.
Claims37
A method for producing a donor Orthomyxovirus for making reassortant viruses, comprising:
- (a) providing a culture of a continuous cell line of monkey kidney cells;
- (b) growing said cell for more than one generation in protein-free medium to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture of step (b) with a virus from the family of Orthomyxovirus;
- (d) selecting for ail Orthomyxovirus strain that exhibits a desired phenotype;
- and
- (e) isolating said donor Orthomyxovirus from step (d).
3 more elements
A method of preventing or treating influenza infection, comprising:
- administering to a subject a vaccine obtainable by
- providing a culture of a continuous cell line of monkey kidney cells;
- growing said cells for more than one generation in protein-free media to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- infecting said culture from the growing step with a virus from the family Orthomyxoviridae;
- incubating said cell culture infected with said virus to propagate virus antigen into said medium;
- harvesting said virus antigen from said culture;
- and
- preparing a vaccine with said harvested virus antigen.
5 more elements
A method for producing a reassortant Orthomyxovirus, comprising:
- (a) providing a culture of a continuous cell line of monkey kidney cells;
- (b) growing said cells for more than one generation in protein-free medium to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said cells with a first Orthomyxovirus having a high-yield phenotype, and a second orthomyxovirus having at least one antigenic determinant;
- (d) incubating said cell of step (b) to propagate said viruses and reassortants of said viruses,
- (e) selecting from said infected culture of step (d) a reassortant virus that comprises the high-yield phenotype from said first Orthomyxovirus and at least one antigenic determinant from said second Orthomyxovirus.
2 more elements
A method fox producing a reassortant Orthomyxovirus, comprising:
- (a) providing a culture of a continuous cell line of monkey kidney cells;
- (b) growing said cells for more than one generation in protein-free medium to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said cells with a first Orthomyxovirus having an attenuated virulence phenotype, and a second orthomyxovirus having at least one antigenic determinant;
- (d) incubating said cells of step (b) to propagate said viruses and reassortants of said viruses,
- (e) selecting from said infected culture of step (d) a reassortant virus that comprises said attenuated virulence phenotype from said first Orthomyxovirus and at least one antigenic determinant from said second Orthomyxovirus.
2 more elements
A method for producing viral antigens acceptable for human administration, comprising:
- (a) providing a culture of a continuous cell line of monkey kidney cells;
- (b) growing said cells for more than one generation in protein-free medium to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture of step (b) with a virus from the family Orthomyxoviridae;
- and
- (d) incubating said cell culture infected with said virus to propagate said virus into said medium to produce said viral antigens.
2 more elements
A method for producing influenza virus vaccine acceptable for human administration, comprising:
- (a) providing a culture of a continuous cell line of monkey kidney cells;
- (b) growing sail cells for more than one generation in protein-free media to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture of step (b) with a virus from the family Orthomyxoviridae;
- (d) incubating said cell culture infected with said virus to propagate virus antigen into said medium;
- (e) harvesting said virus antigen from said culture;
- and
- (f) preparing a vaccine with said harvested virus antigen.
4 more elements
A method for propagating influenza virus, comprising:
- (a) providing a culture of a continuous cell line of monkey kidney cells;
- (b) growing said cells for more than one generation in protein-free medium to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture of step (b) with a virus from the family Orthomyxoviridae that has been isolated directly from an infected mammal;
- and
- (d) incubating said cell culture infected with said virus to propagate said virus.
2 more elements
A method for producing Influenza virus antigen with mammalian Influenza virus having increased infectivity, wherein the antigen is suitable for human administration, comprising the steps of:
- (a) providing a culture comprising a continuous cell line of monkey kidney cells;
- (b) growing said cells in protein-free medium for more than one generation to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture with a mammalian strain of an Influenza virus that has a modified cleavage site in its hemagglutinin, wherein said modified cleavage site increases the susceptibility of said hemagglutinin to a substance that augments the activation of said virus;
- and
- (d) incubating said cell culture of step (c) to propagate said virus and thereby produce said Influenza virus antigen that is suitable for human administration.
2 more elements
A method for producing Influenza virus vaccine suitable for human administration, comprising the steps of:
- (a) providing a culture comprising a continuous cell line of monkey kidney cells;
- (b) growing said cells in protein-free medium for more than one generation to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture with a mammalian strain of an Influenza virus that has a modified cleavage site in its hemagglutinin, wherein said modified cleavage site increases the susceptibility of said hemagglutinin to a substance that augments the activation of said virus;
- (d) incubating said cell culture of step (c) to propagate said virus;
- (e) harvesting said virus;
- and
- (f) preparing a vaccine with the harvested virus, wherein the vaccine is suitable for human administration.
4 more elements
A method for the treatment of Influenza virus infection or for the prevention of Influenza virus infection comprising the step of administering to a human a vaccine obtainable by a method comprising the steps of:
- (a) providing a culture comprising a continuous cell line of monkey kidney cells;
- (b) growing said cells in protein-free medium for more than one generation to ensure (i) adaptation of the cells to protein-free media and (ii) virus production capability;
- (c) infecting said culture with a mammalian strain of an Influenza virus that has a modified cleavage site in its hemagglutinin, wherein said modified cleavage site increases the susceptibility of said hemagglutinin to a substance that augments the activation of said virus;
- (d) incubating said cell culture of step (c) to propagate said virus;
- (e) harvesting said virus;
- and
- (f) preparing a vaccine with the harvested virus, wherein the vaccine is suitable for human administration.
4 more elements
Dependencies are inferred from the claim text.