Automation of cell line development |
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Authors: | Kristina Lindgren Andréa Salmén Mats Lundgren Lovisa Bylund Åsa Ebler Eric Fäldt Lina Sörvik Christel Fenge Ulrica Skoging-Nyberg |
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Institution: | BioProcess R&D, AstraZeneca (now Recipharm Biologics AB), Gärtunavägen 10, 152 57 Södertälje, Sweden |
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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 μM, 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 μM MSX, fewer
low producing transfectants were obtained, compared to cell lines selected with 56 μM 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.
A. Salmén and K. Lindgren contributed equally to the work. |
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Keywords: | Chinese hamster ovary cells Automation Cello robotic system Cell line development Glutamine synthetase |
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