| Abstrakt: |
Microbioreactors have been proven to be a useful tool in high‐throughput applications, such as clone screening, synthetic library testing, and media optimization. Most were designed for low cell density applications, where the optical density of the cultures typically does not exceed an OD600 of 10. In microbial applications, where protein is to be expressed, such a scale is not sufficient to produce material for extensive target molecule testing. Here, we present a method for growing high‐cell density Escherichia coli cultures in milliliter‐scale bioreactors, to produce milligram quantities of target protein. We used a micro‐Matrix system with a starting volume of 3 ml per culture. A combination of defined medium, a fed‐batch feeding strategy at low temperature, and an advanced self‐adapting control algorithm achieved up to 0.7 g of wet cell weight (WCW) in a 5.7 ml final culture volume, which corresponds to 123 g/L WCW. This translates to an estimated protein yield of 1150 mg of target protein per liter final volume. [ABSTRACT FROM AUTHOR] |
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