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Use of at-line and in-situ near-infrared spectroscopy to monitor biomass in an industrial fed-batch Escherichia coli process

Authors

  • S. Alison Arnold,

    Corresponding author
    1. Strathclyde Fermentation Centre, Department of Bioscience and Biotechnology, University of Strathclyde, 204 George Street, Glasgow G1 1XW, United Kingdom; telephone: +44 (0)141-548-3388; fax: +44 (0)141-553-4124
    • Strathclyde Fermentation Centre, Department of Bioscience and Biotechnology, University of Strathclyde, 204 George Street, Glasgow G1 1XW, United Kingdom; telephone: +44 (0)141-548-3388; fax: +44 (0)141-553-4124
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  • Rumpai Gaensakoo,

    1. Strathclyde Fermentation Centre, Department of Bioscience and Biotechnology, University of Strathclyde, 204 George Street, Glasgow G1 1XW, United Kingdom; telephone: +44 (0)141-548-3388; fax: +44 (0)141-553-4124
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  • Linda M. Harvey,

    1. Strathclyde Fermentation Centre, Department of Bioscience and Biotechnology, University of Strathclyde, 204 George Street, Glasgow G1 1XW, United Kingdom; telephone: +44 (0)141-548-3388; fax: +44 (0)141-553-4124
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  • Brian McNeil

    1. Strathclyde Fermentation Centre, Department of Bioscience and Biotechnology, University of Strathclyde, 204 George Street, Glasgow G1 1XW, United Kingdom; telephone: +44 (0)141-548-3388; fax: +44 (0)141-553-4124
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Abstract

One of the key goals in bioprocess monitoring is to achieve real-time knowledge of conditions within the bioreactor, i.e., in-situ. Near-infrared spectroscopy (NIRS), with its ability to carry out multi-analyte quantification rapidly with little sample presentation, is potentially applicable in this role. In the present study, the application of NIRS to a complex, fed-batch industrial E. coli (RV308/PHKY531) process was investigated. This process undergoes a series of temperature changes and is vigorously agitated and aerated. These conditions can pose added challenges to in-situ NIRS. Using the measurement of a key analyte (biomass) as an illustration, the details of the relationship between the at-line and in-situ use of NIRS are considered from the viewpoint of both theory and practical application. This study shows that NIRS can be used both at-line and in-situ in order to achieve good predictive models for biomass. There are particular challenges imposed by in-situ operation (loss of wavelength regions and noise) which meant the need for signal optimisation studies. This showed that whilst the at-line modelling process may provide some useful information for the in-situ process, there were distinct differences. This study shows that the in-situ use of NIRS in a highly challenging matrix (similar to those encountered in current industrial practice) is possible, and thus extends previous works in the area. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 405–413, 2002.

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