Abstract
A quantum dot method for highly efficient labelling of single adenoviral particles is developed. The technique has no impact on viral fitness and allows the imaging and tracking of virus binding and internalisation events using a variety of techniques including imaging cytometry and confocal microscopy. The method is applied to characterise the tropism of different adenoviral vectors.
Highlights
Genetic modification of adenoviral capsid proteins is a commonly used strategy for broadening the tissue tropism of adenoviral gene therapy vectors and increasing their therapeutic potential
We have utilised streptavidin-conjugated quantum dot technology to detect the binding of biotinylated human adenovirus and adenoviral gene therapy vectors to target cells
In order to determine the level of biotinylation compatible with retention of virus viability, a replication deficient Ad vector expressing eGFP (AdGFP)[55] which had been purified [56] was chemically biotinylated by incubation with increasing concentrations of Nhydroxysuccinimido-biotin (NHS-biotin) (10, 100 and 1000 μg/ml for 4 hours, room temperature) and free NHS-biotin removed by dialysis
Summary
Genetic modification of adenoviral capsid proteins is a commonly used strategy for broadening the tissue tropism of adenoviral gene therapy vectors and increasing their therapeutic potential.
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