title: Automation of cell line development.
abstract: An automated platform for development of high producing cell lines for biopharmaceutical production has been established in order to increase throughput and reduce development costs. The concept is based on the Cello robotic system (The Automation Partnership) and covers screening for colonies and expansion of static cultures. In this study, the glutamine synthetase expression system (Lonza Biologics) for production of therapeutic monoclonal antibodies in Chinese hamster ovary cells was used for evaluation of the automation approach. It is shown that the automated procedure is capable of producing cell lines of equal quality to the traditionally generated cell lines in terms of colony detection following transfection and distribution of IgG titer in the screening steps. In a generic fed-batch evaluation in stirred tank bioreactors, IgG titers of 4.7 and 5.0 g/L were obtained for best expressing cell lines. We have estimated that the number of completed cell line development projects can be increased up to three times using the automated process without increasing manual workload, compared to the manual process. Correlation between IgG titers obtained in early screens and titers achieved in fed-batch cultures in shake flasks was found to be poor. This further implies the benefits of utilizing a high throughput system capable of screening and expanding a high number of transfectants. Two concentrations, 56 and 75 muM, of selection agent, methionine sulphoximine (MSX), were applied to evaluate the impact on the number of colonies obtained post transfection. When applying selection medium containing 75 muM MSX, fewer low producing transfectants were obtained, compared to cell lines selected with 56 muM MSX, but an equal number of high producing cell lines were found. By using the higher MSX concentration, the number of cell line development projects run in parallel could be increased and thereby increasing the overall capacity of the automated platform process.
authors.affiliation: BioProcess R&D, AstraZeneca (now Recipharm Biologics AB), Gärtunavägen 10, 152 57, Södertälje, Sweden.
authors.forename: Kristina
authors.initials: K
authors.lastname: Lindgren
authors.forename: Andréa
authors.initials: A
authors.lastname: Salmén
authors.forename: Mats
authors.initials: M
authors.lastname: Lundgren
authors.forename: Lovisa
authors.initials: L
authors.lastname: Bylund
authors.forename: Asa
authors.initials: A
authors.lastname: Ebler
authors.forename: Eric
authors.initials: E
authors.lastname: Fäldt
authors.forename: Lina
authors.initials: L
authors.lastname: Sörvik
authors.forename: Christel
authors.initials: C
authors.lastname: Fenge
authors.forename: Ulrica
authors.initials: U
authors.lastname: Skoging-Nyberg