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

Int J Biol Macromol. 2006 Aug 15;39(1-3):77-82. doi: 10.1016/j.ijbiomac.2006.02.003. Epub 2006 Mar 10.

Abstract

Recombinant adenoviral vectors encoding human HDL-cholesterol receptor SR-BI (Ad/hSR-BI) or beta-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 degrees 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Adenoviridae / isolation & purification*
  • Animals
  • Chromatography, Ion Exchange
  • Cryopreservation
  • Genetic Vectors / genetics
  • Genetic Vectors / isolation & purification*
  • Humans
  • Polymerase Chain Reaction
  • Rabbits
  • Scavenger Receptors, Class B / genetics
  • Transduction, Genetic

Substances

  • Scavenger Receptors, Class B