Patent ID: 8642314
Filing Date: 2014-02-04
Classification: C07K,C12N

Abstract:
1. A method for the production of a recombinant parvoviral virion, which comprises culturing an insect cell comprising one or more nucleic acid constructs which comprise: (a) a nucleotide sequence comprising a transgene that is flanked by at least one parvoviral inverted terminal repeat (ITR) nucleotide sequence; (b) a first expression cassette comprising a nucleotide sequence encoding at least one of parvoviral Rep proteins Rep78 and Rep68, which nucleotide sequence is operably linked to a first promoter that is capable of driving expression of the at least one of Rep78 and Rep68 proteins in the insect cell; and (c) a second expression cassette comprising a nucleotide sequence encoding a parvoviral capsid protein which is operably linked to a second promoter that is capable of driving expression of the capsid protein in the insect cell; under conditions conducive to the expression of the Rep protein and the capsid protein; wherein (i) the first and second expression cassettes are present on a single nucleic acid construct and are present in equimolar amounts in the insect cell, (ii) the ratio of the Rep78 and/or Rep68 protein expression to the capsid protein expression is regulated by one or more of the following:  resulting in expression of the Rep78 and/or Rep68 protein that is greater than expression of the capsid protein.