Application of spectral deconvolution and inverse mechanistic modelling as a tool for root cause investigation in protein chromatography

Journal of Chromatography. a
Nina BrestrichJ Hubbuch

Abstract

In chromatographic protein purification, process variations, aging of columns, or processing errors can lead to deviations of the expected elution behavior of product and contaminants and can result in a decreased pool purity or yield. A different elution behavior of all or several involved species leads to a deviating chromatogram. The causes for deviations are however hard to identify by visual inspection and complicate the correction of a problem in the next cycle or batch. To overcome this issue, a tool for root cause investigation in protein chromatography was developed. The tool combines a spectral deconvolution with inverse mechanistic modelling. Mid-UV spectral data and Partial Least Squares Regression were first applied to deconvolute peaks to obtain the individual elution profiles of co-eluting proteins. The individual elution profiles were subsequently used to identify errors in process parameters by curve fitting to a mechanistic chromatography model. The functionality of the tool for root cause investigation was successfully demonstrated in a model protein study with lysozyme, cytochrome c, and ribonuclease A. Deviating chromatograms were generated by deliberately caused errors in the process parameters flow rate a...Continue Reading

References

Oct 11, 2005·Journal of Chromatography. a·Niklas JakobssonBernt Nilsson
Nov 28, 2006·Journal of Chromatography. a·Niklas JakobssonBernt Nilsson
Mar 19, 2011·Biotechnology Journal·Jarka GlasseyCarl-Fredrik Mandenius
Apr 16, 2014·Trends in Biotechnology·Sarah M MercierMathieu Streefland
Nov 25, 2014·Journal of Pharmaceutical Sciences·Konstantin B Konstantinov, Charles L Cooney

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Citations

Feb 3, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Egbert Müller, Judith Vajda
Nov 14, 2016·Journal of Chromatography. a·Johannes WinderlJürgen Hubbuch
May 21, 2019·Preparative Biochemistry & Biotechnology·Lalita K Shekhawat, Anurag S Rathore
Jun 10, 2020·Annual Review of Chemical and Biomolecular Engineering·Vijesh Kumar, Abraham M Lenhoff
Feb 2, 2021·Biotechnology and Bioengineering·Jon CoffmanJoseph Shultz

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