Biology Reference
In-Depth Information
Chapter 16
Production of Chimeric Adenovirus
Marta Miralles, Marc Garcia, Marcos Tejero, Assumpció Bosch,
and Miguel Chillón
Abstract
The use of chimeric pseudotyped vectors is a common way to modify the adenoviral tropism by replacing
the fiber protein. In this chapter the procedure to generate a chimeric adenovirus pre-stock from a plasmid
containing the adenoviral genome is described. Also, the chimeric adenovirus replicative cycle to increase
the yield in further productions is determined. Finally, two different protocols, in culture plates and in
suspension cultures, to produce the virus at large scale are also detailed.
Key words
Adenoviral vector, Chimeric vectors, Adenovirus production
1
Introduction
Although more than 50 serotypes of HAdVs have been described
[
1
], group C (HAdV-2 and HAdV-5) is still the most studied
group [
2
]. Thus, vectors based on human adenovirus serotype 5
(HAdV-5) have been widely employed as vehicles for many strate-
gies because they have several advantages over other gene delivery
systems. These vectors can be amplified to very high titers, which
is critical for in vivo assays and clinical applications [
3
]. Moreover,
HAdV-5 vectors are relatively safe because HAdV genome does
not integrate into the host genome [
4
,
5
]. For these reasons,
HAdV-5 and chimeric HAdV-5 derived vectors have been prefer-
ably used among other serotypes.
The term “chimeric virus” is generally used for a recombinant
virus generated by the combination of two different viral genomes.
Thus, the new chimeric virus may display a combination of the
biological properties of both parent viruses. Usually, chimeric ade-
noviruses have been generated by fiber replacement, involving the
HAdV-5 backbone and the fiber protein of a different serotype to
thus modify the transduction efficiency and/or the transduction
