MODIFIED VACCINIA ANKARA VIRUS VARIANT AND CULTIVATION METHOD

• US 20110217757A1
• Filed: 05/12/2011
• Published: 09/08/2011
• Est. Priority Date: 11/23/2000
• Status: Active Grant

• Current Assignee: Bavarian Nordic A/S
• Original Assignee: Bavarian Nordic A/S
• Inventors: MEISINGER-HENSCHEL, CHRISTINE, CHAPLIN, PAUL, HELLER, KARL, RATHE, INGMAR, FELDER, EVA, HOWLEY, PAUL
• Granted Patent: US 8,236,560 B2
• Time in Patent Office: Days
• US Class Current: 435/239
• CPC Class Codes: A61K 2039/5254 avirulent or attenuated
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/285 Vaccinia virus or variola virus
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/24134 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2710/24143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2710/24151 Methods of production or purification of viral material
• CPC Class Codes: C12N 2740/16322 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The present invention provides an attenuated virus, which is derived from Modified Vaccinia Ankara virus and characterized by the loss of its capability to reproductively replicate in human cell lines. It further describes recombinant viruses derived from this virus and the use of the virus, or its recombinants, as a medicament or vaccine. A method is provided for inducing an immune response in individuals who may be immune-compromised, receiving antiviral therapy, or have a pre-existing immunity to the vaccine virus. In addition, a method is provided for the administration of a therapeutically effective amount of the virus, or its recombinants, in a vaccinia virus prime/vaccinia virus boost inoculation regimen. The present invention relates to a method of virus amplification in primary cells which are cultivated in a serum free medium. Viruses produced by this method are advantageously free of any infectious agents comprised in animal sera.

Claims

1-68. -68. (canceled)

69. A method for the re-derivation of a virus comprising:

a) infecting chicken embryo fibroblast cells, which have not previously been cultured in a medium containing serum, with a modified vaccinia Ankara virus;

b) cultivating the infected cells in a serum free medium comprising epidermal growth factor at a concentration of 1-50 ng/ml; and

c) isolating the virus produced by the infected cells.

70. The method of claim 69, wherein the serum free medium comprises a fibronectin.

71. The method of claim 69, wherein the epidermal growth factor is human epidermal growth factor.

72. The method of claim 71, wherein the epidermal growth factor is recombinant human epidermal growth factor at a concentration of 5-50 ng/ml.

73. The method of claim 72, wherein the epidermal growth factor is recombinant human epidermal growth factor at a concentration of 5-20 ng/ml.

74. The method of claim 73, wherein the serum free medium comprises human plasma fibronectin at a concentration of 1-10 μ

g/cm².

75. The method of claim 73, wherein the serum free medium comprises a microbial extract, a plant extract, or an extract from a non-mammalian animal.

76. The method of claim 73, wherein the serum free medium comprises asparagine in the range of 0.8-1.8 mM.

77. The method of claim 73, wherein the serum free medium comprises glutamine in the range of 1-5 mM.

78. The method of claim 69, wherein the infected cells are cultivated as adherent cells attached to a solid support.

79. The method of claim 73, wherein the infected cells are cultivated as adherent cells attached to a solid support.

80. The method of claim 69, wherein the infected cells are cultivated in suspension culture.

81. The method of claim 73, wherein the infected cells are cultivated in suspension culture.

82. The method of claim 69, wherein the infected cells are passaged only one time in the serum free medium.

83. The method of claim 73, wherein the infected cells are passaged only one time in the serum free medium.

84. The method of claim 69, wherein the infected cells are passaged 1-8 times in the serum free medium.

85. The method of claim 69, wherein the infected cells are passaged 3-8 times in the serum free medium.

86. The method of claim 73, wherein the infected cells are passaged 3-8 times in the serum free medium.

87. The method of claim 69, wherein the infected cells are passaged 4-6 times in the serum free medium.

88. The method of claim 73, wherein the infected cells are passaged 4-6 times in the serum free medium.