Applications of PAT-process analytical technology in recombinant protein processes with Escherichia coli

Monitoring of bioprocesses and thus observation and identification of such processes is one of the main aims of bioprocess engineering. It is of vital importance in bioprocess development to improve the overall productivity by avoiding unintentional limitations to ensure not only optimal process conditions but also the observation of established production processes. Furthermore, reproducibility needs to be improved and final product quality and quantity be guaranteed. Therefore, an advanced monitoring and control system has been developed, which is based on different in-line, on-line and at-line measurements for substrates and products. Observation of cell viability applying in-line radio frequency impedance measurement and on-line determination of intracellular recombinant target protein using the reporter protein T-Sapphire GFP based on in-line fluorescence measurement show the ability for the detection of critical process states. In this way, the possibility for the on-line recognition of optimal harvest times arises and disturbances in the scheduled process route can be perceived.