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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 toolbox for impurity pattern monitoring during inclusion body processing
Vignesh Rajamanickam1,2, Britta Eggenreich1, Christoph Herwig1,2 and Oliver Spadiut1,2*
1Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Vienna, Austria
Escherichia coli is a well-studied recombinant host organism extensively used for recombinant protein production. Albeit
E. coli is attributed with high product titres and growth on inexpensive media, overexpression of heterologous proteins
often leads to accumulation of target protein in water-insoluble, misfolded aggregates called inclusion bodies (IBs). In practice,
IBs are washed, solubilized and refolded to recover the target protein in its active form. Empirical complex washing and
solubilization protocols are diverse and product specific and often render poor product quality with different impurities, which
negatively impact subsequent refolding. Thus, fast and precise monitoring tools are needed to follow the impurity pattern along
IB processing to judge the efficiency of each Unit Operation. However, such reliable monitoring tools are still scarce to date.
In this study, we developed a novel toolbox using UV chromatogram fingerprints and chemometric techniques to monitor
impurity pattern in IB washing and solubilization. Furthermore, we were not only able to monitor the process but also identify
the optimal time point of transfer from one Unit Operation to the next. We are convinced that this toolbox will not only
facilitate DSP monitoring, but will also allow enhanced process control in the future. The different unit operations where the
novel toolbox was implemented is shown in Figure 1.
Figure 1. Implementation of novel toolbox for monitoring different unit operations involved in inclusion body processing
Keywords: process analytical technology, inclusion body processing, HPLC, process monitoring, Escherichia coli,
chemometrics, chromatography
Biography
Vignesh Rajamanickam procured his Master of Science in pharmaceutical biotechnology from Hamburg University of applied sciences, Germany and, Bachelor
of Technology in Biotechnology from Anna University, India. He started his PhD on March 2014 in Biochemical engineering from Vienna University of Technology
(VUT), Austria. Currently, he is working as a project assistant for developing a novel PAT tool for bioprocess monitoring and control in Christian Doppler laboratory
for mechanistic and physiological methods for improved bioprocesses, VUT, Austria.
Vignesh Rajamanickam et al., J Chromatogr Sep Tech 2016, 7:5(Suppl)