Adenoviral Polypeptide IX Increases Adenoviral Gene Therapy Vector Productivity and Infectivity

An adenoviral gene therapy vector having a genome comprising an adenoviral packaging signal which does not have a cre/lox site flanking it, produced in a producer cell which expresses adenovirus protein IX even when not infected or transduced by an adenovirus.

An adenoviral gene therapy vector comprising adenovirus protein IX and an expressible transgene, the adenoviral gene therapy vector having a genome which does not have an expressed ply gene.

A cell which expresses adenovirus protein IX even when not infected or transduced by an adenovirus, and which expresses adenovirus protein IX in a greater amount than does a similar cell that has been infected with wild-type adenovirus at a Multiplicity of Infection of not more than 1.

A suspension-cultured cell that expresses adenovirus protein IX even when not infected or transduced by an adenovirus.

The suspension-cultured cell of claim 69, where the cell is a human embryonic kidney cell.

A method for manufacturing a pix-deleted adenoviral gene therapy vector in suspension cell culture, comprising:

• culturing in suspension cell culture a producer cell which expresses adenoviral protein IX even if not infected or transduced by adenovirus, transforming the cell with a pix-deleted adenoviral gene therapy vector genome, culturing the cell in suspension while the cell produces a pix-deleted adenoviral gene therapy vector, and then harvesting adenoviral gene therapy vector comprising adenovirus protein IX and a therapeutic transgene.

An adenoviral gene therapy vector manufacturing process comprising obtaining human cells, transducing or transfecting those cells with expressible nucleic acid coding for adenovirus and with expressible nucleic acid coding for adenovirus protein IX, and with nucleic acid coding for a transgene, and then culturing the cells in suspension culture to produce adenoviral gene therapy vector comprising adenovirus protein IX and the transgene, and then harvesting the adenoviral gene therapy vector comprising adenovirus protein IX and the transgene.

A virus manufacturing process comprising obtaining human producer cells that express adenovirus protein IX, and then transducing or transfecting those cells with nucleic acid coding for an adenoviral gene therapy vector, and then culturing the cells in suspension culture to produce adenovirus protein IX and the adenoviral gene therapy vector, and then harvesting adenoviral gene therapy vector comprising adenovirus protein IX.

A manufacturing method for increasing the yield of adenoviral gene therapy vector, comprising manufacturing an adenoviral gene therapy vector in a producer cell that expresses adenovirus protein IX even when not infected or transduced by adenovirus and then harvesting adenoviral gene therapy vector, whereby the ratio of infective adenoviral gene therapy vector produced to viral genomes produced is at least about 20% greater than the ratio obtained when producing the same adenoviral gene therapy vector in a producer cell that does not expresses adenoviral Protein IX when not infected or transduced by adenovirus.