Method for high-efficiency fed-batch cultures of recombinant Escherichia coli
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Updated: Oct 21, 2026

Rapid Protocol for Preparation of Electrocompetent Escherichia coli and Vibrio cholerae
Published on: October 8, 2013
Thomas P Caldwell1, Benjamin F Synoground1, Sarah W Harcum1
1Department of Bioengineering, 301 Rhodes Research Center, Clemson University, Clemson, SC, United States.
Abstract:
Fed-batch processes are commonly used in industry to obtain sufficient biomass and associated recombinant protein or plasmids. In research laboratories, it is more common to use batch cultures, as the setup of fed-batch processes can be challenging. This method outlines a robust and reliable means to generate Escherichia coli biomass in a minimum amount of fermentation time using a standardized fed-batch process. Final cell densities can reach over 50g dry cell weight per liter (gdcw/L) depending on the strain. This method uses a predefined exponential feeding strategy and conservative induction protocol to achieve these targets without multiple trial and error studies. If desired, productivity can be optimized by balancing the induction time and feed rates. This method utilizes cost-efficient defined media, minimizes process control complexity, and potentially aids downstream purification.
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