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Chromatography 2016
September 21-23, 2016
Volume 7, Issue 5(Suppl) J Chromatogr Sep Tech 2016
ISSN: 2157-7064 JCGST, an open access journal
conferenceseries .com
September 21-23, 2016 Amsterdam, Netherlands
World Congress on Chromatography
A novel and fast approach to monitor cell disruption efficiency
Britta Eggenreich, Vignesh Rajamanickam, Christoph Herwig and Oliver Spadiut
Vienna University of Technology, Austria
The bacterium Escherichia coli, is a well-studied recombinant host organismwith a plethora of applications in biotechnology.
High valuable biopharmaceuticals, such as recombinant enzymes, antibody fragments and growth factors, are currently
being produced in E. coli. These molecules are usually produced intra-cellularly which is why cell disruption is required as
the first step in the downstream process. For that purpose, high pressure homogenization is the system of choice since it is
scalable and can be run in continuous mode. However, it is crucial to determine cell disruption efficiency to: Avoid product
loss in intact cells, but also to avoid unnecessary long disruption cycles and thus harm the product. Usually, cell disruption
efficiency is evaluated either by determination of colony forming units (CFUs) or photometric measurements of nucleic acids
and protein content in the lysate. However, these methods are both characterized by disadvantages, as CFUs can only be
counted on the next day, resulting in great time delay, and photometric measurements are affected by matrix effects. In this
study, we implemented a novel online tool based on UV chromatogram fingerprints and chemometric techniques to monitor
cell disruption efficiency. We used: 1) Measurement of the total protein content in the supernatant, 2) determination of CFUs
and 3) flow cytometry as reference analytics to validate this novel tool. Finally, we performed a design of experiments study,
where we changed the factors concentration of biomass per ml buffer, number of homogenization cycles and pressure during
homogenization to analyze and optimize the unit operation high pressure homogenization for a recombinant E. coli strain
producing a highly valuable antibody fragment. Summarizing, we could nicely demonstrate the power of the novel online tool,
which will certainly facilitate the evaluation of this crucial unit operation in the future.
Biography
Britta Eggenreich has finished her studies of Pharmaceutical Science at the University of Vienna in the year 2011. She has worked for two years as a Pharmacist
in Vienna. In 2014, she started her Doctoral thesis in the group of Biochemical Engineering at Vienna University of Technology. Currently, her main focus is the
downstream development for inclusion bodies of a novel antibody fragment produced in E. coli.
Britta Eggenreich et al., J Chromatogr Sep Tech 2016, 7:5(Suppl)