Production of recombinant proteins as inclusion bodies (IBs) in E. Coli requires the development of a process to separate the IBs from soluble and insoluble host contaminants. Separation of the more dense IBs is typically achieved with continuous centrifugation.  Historically, the main challenge of IB recovery process development has been predicting centrifuge performance across scales and centrifuge types. In this presentation, we report the development of a robust and scalable process for the recovery of two different types of IBs from E. Coli fermentations. Lab and pilot scale centrifugation conditions were selected using the concept of equivalent settling area to match production scale centrifuges and the resulting data was leveraged to rapidly scale up this process from 1 L to 100 L and finally to 4500 L fermentations. While 1 L scale recoveries did not adequately represent the 100 L pilot recovery, pilot recoveries were acceptably predictive of the performance of the 4500 L centrifuge. In order to achieve this similarity, differences in the discharge mechanisms between scales must be taken into account, making necessary adjustments to the pilot centrifuge operating parameters.  The 100 L scale-down model was used to characterize the recovery processes in preparation for process validation.

Development, scale-up, and characterization of inclusion body recovery processes

247th National Meeting (2014)

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