Abstract
BackgroundCell disruption is a key unit operation to make valuable, intracellular target products accessible for further downstream unit operations. Independent of the applied cell disruption method, each cell disruption process must be evaluated with respect to disruption efficiency and potential product loss. Current state-of-the-art methods, like measuring the total amount of released protein and plating-out assays, are usually time-delayed and involve manual intervention making them error-prone. An automated method to monitor cell disruption efficiency at-line is not available to date.ResultsIn the current study we implemented a methodology, which we had originally developed to monitor E. coli cell integrity during bioreactor cultivations, to automatically monitor and evaluate cell disruption of a recombinant E. coli strain by high-pressure homogenization. We compared our tool with a library of state-of-the-art methods, analyzed the effect of freezing the biomass before high-pressure homogenization and finally investigated this unit operation in more detail by a multivariate approach.ConclusionA combination of HPLC and automated data analysis describes a valuable, novel tool to monitor and evaluate cell disruption processes. Our methodology, which can be used both in upstream (USP) and downstream processing (DSP), describes a valuable tool to evaluate cell disruption processes as it can be implemented at-line, gives results within minutes after sampling and does not need manual intervention.
Highlights
Cell disruption is a key unit operation to make valuable, intracellular target products accessible for further downstream unit operations
State‐of‐the‐art methods compared to HPLC combined with automated data analysis The goal of this work packages (WPs) was to evaluate the applicability and accuracy of our method of HPLC and automated data analysis, which we successfully used in upstream processing [25], to analyze cell disruption efficiency
State‐of‐the‐art methods compared to HPLC combined with automated data analysis In WP1, we compared different methods to evaluate cell disruption efficiency
Summary
Cell disruption is a key unit operation to make valuable, intracellular target products accessible for further downstream unit operations. Independent of the applied cell disruption method, each cell disruption process must be evaluated with respect to disruption efficiency and potential product loss. An automated method to monitor cell disruption efficiency at-line is not available to date. E. coli has become one of the most important host organisms for the recombinant production of biopharmaceuticals. More than 25% of approved biopharmaceuticals are expressed in this organism [3]. There is a variety of different methods available to disrupt E. coli
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have