This Concentration and diafiltration of whole cells protocol is intended for isolating, concentrating and diafiltering recombinant cells from fermentation broth. This process has been repeatedly implemented with consistent success in E. coli and Pichia fermentation operations.
The filtration isolates the bacteria from the broth components with an ultrafiltration membrane (UF) to pass the broth components freely into the permeate and retain the cells. The protocol calls for the cells starting with an OD of 100 or less to be concentrated to 5X prior to starting the diafiltration. After diafiltration the cells are concentrated to a 80% total solids cell paste.
The system uses the OPTISEP 11000 module with RC100 kD UF membrane and 1.5 mm channel height to
concentrate the process stream and then perform a diafiltration. The process volume for the first step is determined by the fermentation volume.
This Concentration and diafiltration of whole cells Case Study describes how a customer evaluated the
SmartFlow® TFF filter technology, comparing it to the cost and efficiency of a large scale centrifuge to concentrate the E. coli from their fermentor. The customers evaluated two conditions, a best case scenario where the process output would be equivalent to the centrifuge option and an enhanced process where the output would result in significant savings of time, material, and provide less volume for the cell lysis step that
follows.
