- Poster Presentation
- Open Access
Automation for higher throughput in protein expression: visions, facts and fictions
© Mahnke et al; licensee BioMed Central Ltd. 2006
- Published: 10 October 2006
- Recombinant Protein
- Automate Process
- Induction Time
- Recombinant Protein Expression
- Cysteine Protease Inhibitor
Down-scaling, parallelization and automation are new trends in the field of recombinant protein expression in the post genomic era [1–3]. During the past years many companies and academic institutions have heavily invested in process and automation technologies. Does this trend keep its promise? Can post genomic protein production issues be overcome with few automated processes?
This abstract wants to highlight two years of experience in running a Protein Production Center in an industrial environment applying the expression systems BEVS, E. coli ( and transient HEK.EBNA). We describe the streamlined and partially automated processes, the automation equipment applied; discuss results from the past two years of experience and strategies to eliminate remaining bottlenecks.
Proteins expressed in a generic way in the E. coli expression system are all his-tagged and often N-terminally fused to thioredoxin or other fusion partners in order to improve solubility. After small scale expression evaluation in 24-deepwell blocks recombinant proteins are produced in 1-L fermenter vessels using fully automated and unattended inductions and temperature shifts. The described method is applicable to all host strains and induction systems, providing the optimal induction time point and harvest for each construct.
Expression of 30 DUB proteins in the BEVS at 10-L scale and processed in a semi-automated mode.
Av. Yield [%]
Small Scale Expression [mg/L]
3 – 140
Large Scale Expression [mg/l]
0 – 270
Large Scale Expression [mg]
0 – 1400
Cross-flow yields [mg]
0 – 825
Pure protein yields [mg]
0 – 103
We have demonstrated that recombinant protein expression can be evaluated, scaled up and proteins be purified in a parallel and (semi-) automated fashion using the BEVS and E. coli expression systems. Pre-requisites are that proteins are appropriately tagged and grouped. However, in order to attain optimal quality and yield of different recombinant proteins by generic processes the influence of protein destabilizing factors needs to be thoroughly understood and managed.
The authors want to thank Rene Amstutz and Hans Kocher for support of this work and Sabine Deutsch, Eveline Eglin, Brendan Kerins, Stefan Dalcher and Sybille Bossart for excellent technical assistance.
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This article is published under license to BioMed Central Ltd.