A label-free methodology for selective protein quantification by means of absorption measurements

Authors

  • Sigrid K. Hansen,

    1. Institute of Engineering in Life Sciences, Section IV: Biomolecular Separation Science, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany, telphone: +49 721 608 42557; fax: +49 721 608 46240
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  • Erik Skibsted,

    1. CMC Project Planning and Management, Novo Nordisk A/S, Denmark
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  • Arne Staby,

    1. CMC Project Planning and Management, Novo Nordisk A/S, Denmark
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  • Jürgen Hubbuch

    Corresponding author
    1. Institute of Engineering in Life Sciences, Section IV: Biomolecular Separation Science, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany, telphone: +49 721 608 42557; fax: +49 721 608 46240
    • Institute of Engineering in Life Sciences, Section IV: Biomolecular Separation Science, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany, telphone: +49 721 608 42557; fax: +49 721 608 46240.
    Search for more papers by this author

Abstract

The application of high throughput experimentation (HTE) in protein purification process development has created an analytical bottleneck. Using a new label-free and non-invasive methodology for analyzing multicomponent protein mixtures by means of spectral measurements, we show that the analytical throughput for selective protein quantification can be increased significantly. An analytical assay based on this new methodology was shown to generate very precise results. Further, the assay was successfully applied as analytics for a resin screening performed in HTE mode. The increase in analytical throughput was obtained without decreasing the level of information when compared to analytical chromatography. This proves its potential as a valuable analytical tool in conjugation with high throughput process development (HTPD). Further, fast selective protein quantification can enhance process control in a commercial production environment and, hence, minimize the need for off-line release analysis. Biotechnol. Bioeng. 2011;108: 2661–2669. © 2011 Wiley Periodicals, Inc.

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