Production and release testing of ovine atadenovirus vectors.
Gene-directed enzyme prodrug therapy (GDEPT) is an emerging approach for the treatment of cancers. A variety of viral vectors have been used to deliver genes that encode the relevant enzymes, and some have been tested in clinical trials. To ensure the potency and efficacy of such vectors and to obtain regulatory approval to administer them to humans, it is necessary to develop a suite of assays that provide quality assurance. New GDEPT vectors based on ovine atadenovirus and Escherichia coli purine nucleoside phosphorylase (PNP) have been developed for first time use in humans in a phase I trial for the treatment of prostate cancer. Here we describe methods for their production together with several quality-control assays. In particular, a functional cell killing assay was devised to measure the potency of PNP-GDEPT vectors, the principles of which could easily be adapted to other systems.
Both
Gerald W
GW
CSIRO Molecular and Health Technologies, North Ryde, New South Wales, Australia.
Cameron
Fiona
F
Collins
Anne
A
Lockett
Linda J
LJ
Shaw
Jan
J
D000257:Adenoviridae Infections / Q000821:virology*; D000818:Animals; D046808:Atadenovirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D002460:Cell Line; D045744:Cell Line, Tumor; D017931:DNA Primers; D015316:Genetic Therapy; D006801:Humans; D008168:Lung; D008297:Male; D016133:Polymerase Chain Reaction; D011471:Prostatic Neoplasms; D011995:Recombination, Genetic; D012756:Sheep / Q000821:virology*; D012757:Sheep Diseases / Q000821:virology*; D014461:Ultracentrifugation
D017931:DNA Primers