title: A novel toolbox for E. coli lysis monitoring.
abstract: The bacterium Escherichia coli is a well-studied recombinant host organism with a plethora of applications in biotechnology. Highly valuable biopharmaceuticals, such as antibody fragments and growth factors, are currently being produced in E. coli. However, the high metabolic burden during recombinant protein production can lead to cell death, consequent lysis, and undesired product loss. Thus, fast and precise analyzers to monitor E. coli bioprocesses and to retrieve key process information, such as the optimal time point of harvest, are needed. However, such reliable monitoring tools are still scarce to date. In this study, we cultivated an E. coli strain producing a recombinant single-chain antibody fragment in the cytoplasm. In bioreactor cultivations, we purposely triggered cell lysis by pH ramps. We developed a novel toolbox using UV chromatograms as fingerprints and chemometric techniques to monitor these lysis events and used flow cytometry (FCM) as reference method to quantify viability offline. Summarizing, we were able to show that a novel toolbox comprising HPLC chromatogram fingerprinting and data science tools allowed the identification of E. coli lysis in a fast and reliable manner. We are convinced that this toolbox will not only facilitate E. coli bioprocess monitoring but will also allow enhanced process control in the future.
authors: Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9/166, 1060, Vienna, Austria.|Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1a, 1060, Vienna, Austria.
authors: Vignesh
authors: V
authors: Rajamanickam
authors: Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9/166, 1060, Vienna, Austria.
authors: David
authors: D
authors: Wurm
authors: Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9/166, 1060, Vienna, Austria.
authors: Christoph
authors: C
authors: Slouka
authors: Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9/166, 1060, Vienna, Austria.|Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1a, 1060, Vienna, Austria.
authors: Christoph
authors: C
authors: Herwig
authors: Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9/166, 1060, Vienna, Austria. oliver.spadiut@tuwien.ac.at.|Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1a, 1060, Vienna, Austria. oliver.spadiut@tuwien.ac.at.
authors: Oliver
authors: http://orcid.org/0000-0003-0916-0644
authors: O
authors: Spadiut
mesh_terms: D000906:Antibodies / Q000235:genetics / Q000378:metabolism; D001431:Bacteriological Techniques / Q000379:methods*; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D002845:Chromatography; D004926:Escherichia coli / Q000502:physiology*; D011994:Recombinant Proteins / Q000302:isolation & purification*
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins
keywords: Bioprocess monitoring; Chromatogram fingerprinting; Data science tools; Escherichia coli; HPLC; Lysis