No coin nor oath required. For personal study only.
Demand for low-cost proteins continues to increase, with generic products bringing renewed interest in secondgeneration processes. E. coli is an upstream host of choice but can express an insoluble inclusion body, leading designers toward alternatives. However, the decision to trade upstream efficiency for downstream simplicity increases overall product cost. Leong and Middelberg demonstrate that simple E. coli expression can be coupled with simple downstream processing to efficiently produce alfa-fetoprotein. Counterintuitively, dissolving the entire cell with urea did not compromise the ability to recover highly purified protein. Purification was necessary before refolding to avoid contaminant-induced aggregation, and problems in using urea were overcome. Could failure to standardize contaminants before refolding be a cause of variability, and development failure, for other products? Could our reluctance to use urea be misplaced? This work steps us toward a new practical flowsheet for inclusion body processing having immediate use for new products and those coming off-patent.
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