Compositions and methods related to an adenoviral trans-complementing cell line
AssigneeIntrogen Therapeutics, Inc.
InventorsPeter Clarke, Shuyuan Zhang, Hai Pham, Joe Senesac
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Abstract
Embodiments of the invention include E1 expressing cell lines that can be used in a variety of methods for production of an E1 defective adenovirus. In certain aspects a cell of the invention can be adapted to various culture conditions, e.g., suspension culture in serum free medium. In a further aspect, the cell lines allow isolation and subculture of E1-deleted recombinant adenoviruses in an environment free of replication competent adenovirus (RCA).
Claims15
An isolated polynucleotide comprising:
- (i) a first DNA segment encoding an adenoviral E1 protein;
- (ii) a second DNA segment encoding an adenoviral protein IX;
- and
- (iii) a heterologous DNA spacer positioned between the first and second DNA segment.
An adenoviral E1 complementing cell line comprising an expression cassette comprising
- (i) a first DNA segment encoding an adenoviral E1 protein;
- (ii) A second DNA segment encoding an adenoviral protein IX;
- and
- (iii) a heterologous DNA spacer positioned between the first and second DNA segment.
(canceled)
A method for producing an E1 deficient recombinant adenoviral vector comprising:
- (a) providing a complementing cell comprising:
- (i) a first DNA segment encoding an adenoviral E1 protein;
- (ii) a second DNA segment encoding an adenoviral protein IX;
- and
- (iii) a heterologous DNA spacer positioned between the first and second DNA segment;
- and
- (b) introducing an E1 deficient adenoviral nucleic acid into the complementing cell;
- (c) culturing the complementing cell;
- and
- (d) harvesting recombinant adenovirus produced from or by the complementing cell.
Dependencies are inferred from the claim text.