- Infection of dividing and non-dividing cells
- transient transgene expression, not integrating into the genome
- 7.5kb DNA uptake capacity
- dsDNA genome
- 120nm capsid size
- Security level S2
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For the generation of adenoviruses we are currently using kits from Clontech. The viral vector is generated by enzyme mediated insertion directly from a purified PCR product of the transgene into the the E1 and E3 deleted Ad5 genome. Transgene expression is driven by the strong CMV promoter.
Viral production is performed by transfection of the linearized Ad-vector into HEK293 cells expressing E1 and E3 genes of the Ad5 genome and several rounds of amplification. Alternatively, existing adenoviruses can simply be used to start the amplification. Purification of properly assembled adenoviral particles is done by caesium chloride density gradient followed by dialysis.
Costs - Adenovirus Production
Adenovirus production using AV-lysates as template
Infection of AV particles into HEK293A cells and purification by cesium chloride density gradient (2 rounds of 20x15cm cell culture plates). scale: app. 1-2ml, 10^10-11 IU/ml
Adenovirus production using plasmid as template
Transfection of AV-vectors into HEK293A cells and purification by cesium chloride density gradient (2 rounds of 20x15cm cell culture plates).
Adenovirus production from supplied cell lysate
Purification by cesium chloride density gradient centrifugation from supplied cell lysate over 2 rounds, scale app. 1-2 ml, 10 ^ 10-12 IU / ml (10-20x15 cm HEK293)