A combination of HPLC and automated data analysis for monitoring the efficiency of high-pressure homogenization

Microbial Cell Factories

Technical Notes

Britta Eggenreich¹  Oliver Spadiut¹  David Johannes Wurm¹  Vignesh Rajamanickam²  Christoph Herwig²  Jens Fricke² 

[1] Research Division Biochemical Engineering, Institute of Chemical, Environmental and Biological Engineering, TU Wien, Gumpendorfer Strasse 1a, 1060, Vienna, Austria;Research Division Biochemical Engineering, Institute of Chemical, Environmental and Biological Engineering, TU Wien, Gumpendorfer Strasse 1a, 1060, Vienna, Austria;Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Institute of Chemical, Environmental and Biological Engineering, TU Wien, Gumpendorferstraße 1a, 1060, Vienna, Austria;

关键词: Cell disruption;    High-pressure homogenization;    E.coli;    HPLC;    Data analysis;   

DOI  :  10.1186/s12934-017-0749-y

 received in 2017-04-27, accepted in 2017-07-24,  发布年份 2017

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© The Author(s) 2017