title: Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale.
abstract: Large-scale vector manufacturing for phase III and beyond has proven to be challenging. Upscaling the process with suspension cells is increasingly feasible, but many viral production applications are still applicable only in adherent settings. Scaling up the adherent system has proven to be troublesome. The iCELLis(®) disposable fixed-bed bioreactors offer a possible option for viral vector manufacturing in large quantities in an adherent environment. In this study, we have optimized adenovirus serotype 5 manufacturing using iCELLis Nano with a cultivation area up to 4 m(2). HEK293 cell cultivation, infection, and harvest of the virus (by lysing the cells inside the bioreactor) proved possible, reaching total yield of up to 1.6×10(14) viral particles (vp)/batch. The iCELLis 500 is designed to satisfy demand for large-scale requirements. Inoculating a large quantity of cell mass into the iCELLis 500 was achieved by first expanding the cell mass in suspension. Upscaling the process into an iCELLis 500/100 m(2) cultivation area cassette was practical and produced up to 6.1×10(15) vp. Flask productivity per cm(2) in iCELLis Nano and iCELLis 500 was in the same range. As a conclusion, we showed for the first time that iCELLis 500 equipment has provided an effective way to manufacture large batches of adenoviral vectors.
authors.affiliation: 1 FKD Therapies, Kuopio, Finland.|2 FinVector Vision Therapies, Kuopio, Finland.|3 A.I. Virtanen Institute for Molecular Sciences, Department of Biotechnology and Molecular Medicine, University of Eastern Finland , Kuopio, Finland.
authors.forename: Hanna P
authors.initials: HP
authors.lastname: Lesch
authors.affiliation: 2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Kati M
authors.initials: KM
authors.lastname: Heikkilä
authors.affiliation: 1 FKD Therapies, Kuopio, Finland.|2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Eevi M
authors.initials: EM
authors.lastname: Lipponen
authors.affiliation: 2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Piia
authors.initials: P
authors.lastname: Valonen
authors.affiliation: 2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Achim
authors.initials: A
authors.lastname: Müller
authors.affiliation: 2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Eva
authors.initials: E
authors.lastname: Räsänen
authors.affiliation: 2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Tarja
authors.initials: T
authors.lastname: Tuunanen
authors.affiliation: 1 FKD Therapies, Kuopio, Finland.|2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Minna M
authors.initials: MM
authors.lastname: Hassinen
authors.affiliation: 1 FKD Therapies, Kuopio, Finland.|2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Nigel
authors.initials: N
authors.lastname: Parker
authors.affiliation: 2 FinVector Vision Therapies, Kuopio, Finland.
authors.forename: Minna
authors.initials: M
authors.lastname: Karhinen
authors.affiliation: 1 FKD Therapies, Kuopio, Finland.|2 FinVector Vision Therapies, Kuopio, Finland.|3 A.I. Virtanen Institute for Molecular Sciences, Department of Biotechnology and Molecular Medicine, University of Eastern Finland , Kuopio, Finland.
authors.forename: Robert
authors.initials: R
authors.lastname: Shaw
authors.affiliation: 3 A.I. Virtanen Institute for Molecular Sciences, Department of Biotechnology and Molecular Medicine, University of Eastern Finland , Kuopio, Finland.|4 Science Service Center and Gene Therapy Unit, Kuopio University Hospital , Kuopio, Finland .
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
mesh_terms: D000256:Adenoviridae / Q000502:physiology*; D019149:Bioreactors; D049109:Cell Proliferation; D003470:Culture Media; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D014776:Virus Cultivation; D014779:Virus Replication
chemical_list: D003470:Culture Media