Biology Reference
In-Depth Information
Chapter 13
Scalable Production of Adenovirus Vectors
Ana Carina Silva, Paulo Fernandes, Marcos F.Q. Sousa,
and Paula M. Alves
Abstract
Recombinant adenoviruses (AdV) are highly efficient at gene transfer for a broad spectrum of cell types
and species. They became one of the vectors of choice for gene delivery and expression of foreign proteins
in gene therapy and vaccination purposes. To meet the need of significant amounts of adenoviral vectors
for preclinical and possibly clinical uses, scalable and reproducible production processes are required.
In this chapter, we review processes used for scalable production of two types of first generation
(E1-deleted) adenoviral vectors (Human and Canine) using stirred tank bioreactors. The production of
adenovirus vectors using either suspension (HEK 293) or anchorage-dependent cells (MDCK-E1) are
described to exemplify scalable production processes with different cell-culture types. The downstream
processes will be covered in the next chapter.
Key words
Adenovirus, Human, Canine, Suspension, Microcarriers, Gene therapy, Vaccination,
Stirred tank bioreactor
1
Introduction
1.1 Adenoviral
Vectors
Viral vectors are currently the most efficient tools for in vivo gene
transfer. Human adenovirus vectors (HAdV) derived from sero-
type 5 are the most characterized viruses among those of the same
family [
1
,
2
]. HAdV has been considered a good candidate for
human gene therapy due to a number of advantages including its
wide cell tropism in quiescent and non-quiescent cells, its inability
to integrate the host genome and its high production titer [
3
].
Currently, adenovirus vectors (AdV) are being tested as subunit
vaccine systems for numerous infectious agents ranging from
malaria to HIV-1 [
4
,
5
]. Additionally, they are being explored as
vaccines against a multitude of tumor-associated antigens. However,
gene transfer efficacy and the clinical use of HAdV can be hampered
by the preexisting humoral and cellular immunity in most humans
[
6
,
7
]. Therefore, alternatives able to bypass some of these clinical
disadvantages, while keeping the numerous advantages associated
