Continuous processing of fusion protein expressed as an Escherichia coli inclusion body |
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Authors: | Morreale Giacomo Lanckriet Heikki Miller Jemma C Middelberg Anton P J |
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Institution: | Department of Chemical Engineering, University of Cambridge, Pembroke Street, CB2 3RA, Cambridge, UK. |
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Abstract: | In this study we develop the components of an integrated process for the continuous extraction and purification of a histidine-tagged fusion protein expressed as an inclusion body in Escherichia coli. Lac21 was selected as a model peptide and was expressed as a fusion to ketosteroid isomerase. A purification strategy was developed on a 1-ml batch column before successful scale-up and transfer to a continuous purification system, having a bed volume of 240 ml. Preliminary experiments proved cleavage of the fusion protein. The use of chemical extraction and continuous chromatography gives a flowsheet far superior to the traditional methods for inclusion body processing. |
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