Replication methods and tools in high-throughput cultivation processes - recognizing potential variations of growth and product formation by on-line monitoring |
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Authors: | Robert Huber Thomas G Palmen Nadine Ryk Anne-Kathrin Hillmer Karina Luft Frank Kensy Jochen Büchs |
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Affiliation: | (1) AVT - Aachener Verfahrenstechnik, Biochemical Engineering, RWTH Aachen University, Worringerweg 1, D-52074 Aachen, Germany;(2) m2p-labs GmbH, Forckenbeckstr. 6, 52074 Aachen, Germany |
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Abstract: | Background High-throughput cultivations in microtiter plates are the method of choice to express proteins from recombinant clone libraries. Such processes typically include several steps, whereby some of them are linked by replication steps: transformation, plating, colony picking, preculture, main culture and induction. In this study, the effects of conventional replication methods and replication tools (8-channel pipette, 96-pin replicators: steel replicator with fixed or spring-loaded pins, plastic replicator with fixed pins) on growth kinetics of Escherichia coli SCS1 pQE-30 pSE111 were observed. Growth was monitored with the BioLector, an on-line monitoring technique for microtiter plates. Furthermore, the influence of these effects on product formation of Escherichia coli pRhotHi-2-EcFbFP was investigated. Finally, a high-throughput cultivation process was simulated with Corynebacterium glutamicum pEKEx2-phoD-GFP, beginning at the colony picking step. |
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