title: Scalable inoculation strategies for microcarrier-based animal cell bioprocesses.
abstract: Scalability is a major demand for high-yield, stable bioprocess systems in animal cell culture-based biopharmaceutical production. Increased yields can be achieved through high-density cell culture, such as in the combination of microcarrier and fluidized bed bioreactor technology. To minimize inocula volume in industrial applications of fluidized bed fermentation systems, it is crucial to increase the bed volume in the reactor during the fermentation process. We tested scale-up strategy for the production of recombinant human arylsulfatase B (ASB) enzyme used in enzyme replacement therapy in patients afflicted with mucopolysaccharidosis type VI (MPS VI). This enzyme was derived from Chinese hamster ovary (CHO) cells cultivated as adherent cell culture on Cytoline macroporous microcarriers (Amersham Biosciences, Uppsala, Sweden) using a Cytopilot Mini fluidized bed bioreactor (FBR; Amersham Biosciences, Vogelbusch, Austria). Both 1:2 expansion (herein referred to as the addition of fresh, not-yet-colonized microcarriers) and 1:6 expansion of the carrier bed were performed successfully; the cells restarted to proliferate for colonizing these newly added carriers; and the stability of the culture was not negatively affected.
authors.affiliation: Institute of Applied Microbiology, Muthgasse 18, A-1190 Vienna, Austria. h9040929@edv1.boku.ac.at
authors.forename: M
authors.initials: M
authors.lastname: Dürrschmid
authors.forename: K
authors.initials: K
authors.lastname: Landauer
authors.forename: G
authors.initials: G
authors.lastname: Simic
authors.forename: G
authors.initials: G
authors.lastname: Blüml
authors.forename: O
authors.initials: O
authors.lastname: Doblhoff-Dier
mesh_terms: D000818:Animals; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D006224:Cricetinae; D020051:N-Acetylgalactosamine-4-Sulfatase; D011994:Recombinant Proteins
chemical_list: D011994:Recombinant Proteins; D020051:N-Acetylgalactosamine-4-Sulfatase