A semi-automated large-scale process for the production of recombinant tagged proteins in the Baculovirus expression system

Schlaeppi, Jean-Marc; Henke, Mario; Mahnke, Marion; Hartmann, Steffen; Schmitz, Rita; Pouliquen, Yann; Kerins, Brendan; Weber, Eric; Kolbinger, Frank; Kocher, Hans P.

doi:10.1016/j.pep.2006.06.021

Cited by 29 publications

(12 citation statements)

References 20 publications

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“…The bag is rocked on a heated platform which ensures that the culture is uniformly warmed to 27°C and that cells are continuously mixed (Singh, 1999). Oxygen transfer rates in the Wave™ bag system (or equivalent) are similar to conventional bioreactors (Weber et al, 2002) and have been incorporated into semi-automated workflows for protein production in insect cells (Schlaeppi et al, 2006). Nevertheless, we and others have found that scale-up from shake flasks to Wave™ bags usually is accompanied by a protein yield penalty although the exact reason is unknown.…”

Section: Scaling Up Production and Labelling With Selenomethioninementioning

confidence: 99%

Recent advances in the production of proteins in insect and mammalian cells for structural biology

Nettleship

Assenberg

Diprose

et al. 2010

Journal of Structural Biology

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Section: Scaling Up Production and Labelling With Selenomethioninementioning

confidence: 99%

Recent advances in the production of proteins in insect and mammalian cells for structural biology

Nettleship

Assenberg

Diprose

et al. 2010

Journal of Structural Biology

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“…Structural genomics initiatives have successfully implemented parallel protein production with E. coli host cells in milligram amounts for crystallization [32]. Recently, a fully integrated process has been described for protein expression and downstream processing using the Baculovirus system [33]. Protein expression is performed in Bio-Wave cell culture bags, cells are washed and lysed by tangential flow filtration and subsequently the lysate is automatically loaded and separated on immobilized metal chelate chromatography columns.…”

Section: Protein Productionmentioning

confidence: 99%

Automated technologies and novel techniques to accelerate protein crystallography for structural genomics

et al. 2008

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The sequence infrastructure that has arisen through large-scale genomic projects dedicated to protein analysis, has provided a wealth of information and brought together scientists and institutions from all over the world. As a consequence, the development of novel technologies and methodologies in proteomics research is helping to unravel the biochemical and physiological mechanisms of complex multivariate diseases at both a functional and molecular level. In the late sixties, when X-ray crystallography had just been established, the idea of determining protein structure on an almost universal basis was akin to an impossible dream or a miracle. Yet only forty years after, automated protein structure determination platforms have been established. The widespread use of robotics in protein crystallography has had a huge impact at every stage of the pipeline from protein cloning, over-expression, purification, crystallization, data collection, structure solution, refinement, validation and data management- all of which have become more or less automated with minimal human intervention necessary. Here, recent advances in protein crystal structure analysis in the context of structural genomics will be discussed. In addition, this review aims to give an overview of recent developments in high throughput instrumentation, and technologies and strategies to accelerate protein structure/function analysis.

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“…In cases where the tendency of these hosts to overglycosylate the target protein poses problems, alternatives are Baculovirus based expression in insect cells, or expression in mammalian TC cells. The insect cell system is under intense scrutiny by academic and industrial laboratories for developing HTP expression screening schemes [42][43][44], but also TC cells, in particular HEK293, appear to be suitable for HTP expression profiling, and some results have been reported ( [45] and references therein), [46]. The consumables costs for HEK293 cells are considerably lower than for insect cells.…”

Section: Expression Systems: Pro-and Eukaryotic Cell Freementioning

confidence: 99%

“…The use of competition ELISA as an online quality control method is particularly widespread in the field of eukaryotic expression [44]. In terms of downstream characterization, ELISA can be used for complex stability and protein integrity assessment.…”

Section: Protein Characterizationmentioning

confidence: 99%

Pipelines, Robots, Crystals and Biology: What Use High Throughput Solving Structures of Challenging Targets?

Kambach

2007

CPPS

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With recent advances in the technology and software underlying crystallographic structure solution, demands on both output and functional significance of X-ray structures are soaring. To achieve the required speed and quality also with ever larger and more difficult targets, combining HTP screening methods (robotics based or not) adopted from structural genomics initiatives with thorough expertise and dedicated characterization effort for each individual target is almost a must. I present concepts, practical considerations, and experiences on implementing an HTP technology platform for structural and functional studies on complexes, membrane proteins and other challenging targets. Emphasis lies on the environment of small academic groups engaged exclusively in hypothesis driven projects focused on specific biological systems. Suitability of given HTP protocols for particular target classes, benchmarking and quality control for procedures, and project management issues at the interface between extensive, broad parameter screening and intensive individual target work required by non-SG amenable targets are discussed.

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A semi-automated large-scale process for the production of recombinant tagged proteins in the Baculovirus expression system

Cited by 29 publications

References 20 publications

Recent advances in the production of proteins in insect and mammalian cells for structural biology

Recent advances in the production of proteins in insect and mammalian cells for structural biology

Automated technologies and novel techniques to accelerate protein crystallography for structural genomics

Pipelines, Robots, Crystals and Biology: What Use High Throughput Solving Structures of Challenging Targets?

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