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Combinatorial analysis of host-leader-Fab can greatly improve integrated process design
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  • Mathias Fink,
  • Clemens Schimek,
  • Monika Cserjan-Puschmann,
  • Daniela Reinisch,
  • Cécile Brocard,
  • Rainer Hahn,
  • Gerald Striedner
Mathias Fink
Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli,University of Natural Resources and Life Sciences
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Clemens Schimek
Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli,University of Natural Resources and Life Sciences , Department of Biotechnology Vienna, AT
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Monika Cserjan-Puschmann
Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli,University of Natural Resources and Life Sciences , Department of Biotechnology Vienna, AT
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Daniela Reinisch
Boehringer Ingelheim RCV GmbH & Co KG, Dr. Boehringer-Gasse 5-11, A-1120 Vienna, Austria
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Cécile Brocard
Boehringer Ingelheim RCV GmbH & Co KG, Dr. Boehringer-Gasse 5-11, A-1120 Vienna, Austria
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Rainer Hahn
Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli, Department of Biotechnology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria
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Gerald Striedner
Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli,University of Natural Resources and Life Sciences , Department of Biotechnology Vienna, AT
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Abstract

Bioprocess development and optimization is a challenging, costly, and time-consuming effort. In this multidisciplinary task, upstream processing (USP) and downstream processing (DSP) are conventionally considered distinct disciplines. This consideration fosters “one-way” optimization without considering interdependencies between unit operations; thus, the full potential of the process chain cannot be achieved. Therefore, it is necessary to fully integrate USP and DSP process development to provide balanced biotechnological production processes. The aim of the present study was to investigate how different host/leader/antigen binding fragment (Fab) combinations in E. coli expression systems influence USP and primary recovery performance and the final product quality. We ran identical fed-batch cultivations with 16 different expression clones to study growth and product formation kinetics, as well as centrifugation efficiency, viscosity, extracellular DNA, and endotoxin content, which are important parameters in DSP. We observed a severe influence on cell growth, product titer, extracellular product, and cell lysis, accompanied by a significant impact on the analyzed parameters of DSP performance. Our results provide the basis for establishing integrated process development considering interdependencies between USP and DSP. These interdependencies need to be understood for rational decision-making and efficient process development.

Peer review status:ACCEPTED

02 Nov 2020Submitted to Biotechnology Journal
11 Nov 2020Submission Checks Completed
11 Nov 2020Assigned to Editor
11 Nov 2020Reviewer(s) Assigned
30 Dec 2020Editorial Decision: Revise Major
14 Jan 20211st Revision Received
14 Jan 2021Assigned to Editor
14 Jan 2021Submission Checks Completed
14 Jan 2021Reviewer(s) Assigned
28 Jan 2021Editorial Decision: Revise Minor
03 Feb 20212nd Revision Received
03 Feb 2021Submission Checks Completed
03 Feb 2021Assigned to Editor
03 Feb 2021Reviewer(s) Assigned
04 Feb 2021Editorial Decision: Accept