Economical parallel protein expression screening and scale-up in Escherichia coli |
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Authors: | Oleg Brodsky Ciarán N Cronin |
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Institution: | (1) Department of Structural Biology, Pfizer Global Research and Development, 10628 Science Center Drive, La Jolla, CA 92121, USA |
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Abstract: | A novel microfermentation and scale-up platform for parallel protein production in Escherichia coli is described. The vertical shaker device Vertiga, which generates low-volume high density (A600 ∼ 20) Escherichia coli cultures in 96-position deep-well plates without auxiliary oxygen supplementation, has been coupled to a new disposable shake
flask design, the Ultra YieldTM flask, that allows for equally high cell culture densities to be obtained. The Ultra YieldTM flask, which accommodates up to 1 l in culture volume, has a baffled base and a more vertical wall construction compared
to traditional shake flask designs. Experimental data is presented demonstrating that the Ultra YieldTM flask generates, on average, an equivalent amount of recombinant protein per unit cell culture density as do traditional
shake flask designs but at a substantially greater amount per unit volume. The combination of Vertiga and the Ultra YieldTM flask provides a convenient and scalable low-cost solution to parallel protein production in Escherichia coli. |
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Keywords: | Ultra YieldTM flask Vertiga Protein expression E coli fermentation structural proteomics High-density fermentation |
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