In vivo application of adenoviral vectors purified by a Taqman Real Time PCR-supported chromatographic protocol

Recombinant adenoviral vectors encoding human HDL-cholesterol receptor SR-BI (Ad/hSR-BI) or β-galactosidase (Ad/lacZ), respectively, were purified using a Source15™Q anion-exchange (AEX) column and quantified using two parallel Taqman Real Time PCR systems with different target sequences. Adenovirus concentrations were ascertained by 260 nm measurements, purity by 260/280 nm ratio and SDS-PAGE. Subsequently, adenoviruses were validated by experimental intravenous application into New Zealand White rabbits. Transgene expression was verified by functional assays determining plasma clearance rate of 3H-HDL-cholesterol, and was not affected by 21-months storage at −80 °C. No alterations of liver enzymes and C-reactive protein (CRP) upon Source15™Q adenovirus treatment could be detected, demonstrating biological safety of our protocol.