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Fast Quantification of Recombinant Protein Inclusion Bodies within Intact Cells by FT-IR Spectroscopy

Authors

  • Sven Gross-Selbeck,

    1. Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria
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    • Contributed equally to this work

  • Gerd Margreiter,

    1. Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria
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    • Contributed equally to this work

  • Christian Obinger,

    1. Department of Chemistry, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria
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  • Karl Bayer

    Corresponding author
    1. Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria
    • Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria
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Abstract

The accomplishment of the quantification of the recombinant protein content of whole bacterial cells by FT-IR spectroscopy by application of chemometrics is shown. Recombinant Escherichia coli cells expressing an inclusion body forming fusion protein were dried on a 96-well silicon plate for the analysis in a high-throughput FT-IR spectrometer. Acquired spectra of additionally conventionally quantified samples were used to establish a multivariate calibration. The obtained method was tested by predicting inclusion body contents of samples not used for the multivariate model. Results from FT-IR spectra coincided well with the data of universalized electrophoresis analysis. Hence FT-IR spectroscopy could prove as a fast and simple alternative to conventional quantification methods.

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