| 摘要 |
A new series of helper cell lines for the complementation, amplification, and controlled attenuation of E1-deleted adenovirus are disclosed in the present invention. These cell lines are advantageous because they can complement adenovirus E1 gene deletions without production of replication competent adenovirus (RCA), thus making them safer for the large-scale production of adenovirus stock for use in human gene therapy trials. A preferred embodiment is an A549E1 cell line that contains only the Ad5 E1 DNA sequences sufficient for complementation of E1-deleted adenoviral vectors without sequences that overlap with the adenovirus vector. In another aspect, the present invention embodies methods for the production of second generation A549-E1 complementing cell lines that, in addition to producing E1, also produce proteins required for further manipulation of adenoviral vectors. A preferred embodiment is an A549E1 cell line with DNA sequences that encode a polypeptide sufficient for packaging attenuation of E1-deleted helper virus, in order to enrich for packaging of mini-adenovirus.
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