title: Stable hybridoma cultivation in a pilot-scale acoustic perfusion system: long-term process performance and effect of recirculation rate.
abstract: Perfusion systems have the possibility to be operated continuously for several months. It is important that the performance of the cell retention device does not limit the operation time of a perfusion process used in the production of active pharmaceutical ingredients. Therefore, the aim of this study was to investigate the reliability and long-term stability of an acoustic perfusion process using the 200 L/d BioSep. As the BioSep is an external device, it is possible that dependent on the recirculation rate nutrient gradients occur in the external loop, which could affect the cell metabolism. Therefore, the effect of possible nutrient gradients on cell metabolism, viability and productivity was studied by varying the recirculation rate. In this study, it is shown that a perfusion process using a pilot-scale acoustic cell-retention device (200 L/d) is reliable and simple to operate, resulting in a stable 75-day cultivation of a hybridoma cell line producing a monoclonal antibody. The recirculation rate had a significant effect on the oxygen concentration in the external loop, with oxygen being depleted within the cell-retention device at recirculation rates below 6 m3/m(reactor)3.d (=600 L/d). The oxygen depletion at low circulation rates correlated with a slightly increased lactate production rate. For all other parameters no effect of the recirculation rate was observed, including cell death measured through the release of lactate dehydrogenase and specific productivity. A maximum specific productivity of 12 pg/cell.d was reached.
authors.affiliation: Department of Agrotechnology and Food Sciences, Food and Bioprocess Engineering Group, Wageningen University, Bomenweg 2, 6703 HD Wageningen, The Netherlands. ciska.dalm@wur.nl
authors.forename: Marcella C F
authors.initials: MC
authors.lastname: Dalm
authors.forename: Menno
authors.initials: M
authors.lastname: Jansen
authors.forename: Timo M P
authors.initials: TM
authors.lastname: Keijzer
authors.forename: Wout M J
authors.initials: WM
authors.lastname: van Grunsven
authors.forename: Arthur
authors.initials: A
authors.lastname: Oudshoorn
authors.forename: Johannes
authors.initials: J
authors.lastname: Tramper
authors.forename: Dirk E
authors.initials: DE
authors.lastname: Martens
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D004526:Efficiency; D005285:Fermentation; D005947:Glucose / Q000032:analysis; D005973:Glutamine / Q000032:analysis; D006825:Hybridomas / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism; D010100:Oxygen / Q000032:analysis; D010477:Perfusion
chemical_list: D000911:Antibodies, Monoclonal; D005973:Glutamine; D019344:Lactic Acid; D005947:Glucose; D010100:Oxygen