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Bioengineering 2018, 5(4), 79; https://doi.org/10.3390/bioengineering5040079

Influence of Incident Wavelength and Detector Material Selection on Fluorescence in the Application of Raman Spectroscopy to a Fungal Fermentation Process

1
Biopharmaceutical Bioprocess Technology Centre, Merz Court, Newcastle University, Newcastle-upon-Tyne NE1 7RU, UK
2
Perceptive Engineering Limited, Vanguard House, Keckwick Lane, Daresbury, Cheshire WA4 4AB, UK
3
School of Science, Engineering & Design, Teeside University, Tees Valley TS1 3BX, UK
4
Control Systems Group, School of Electrical and Electronic Engineering, University of Manchester, Manchester M13 9PL, UK
Current address: Department of Biochemical Engineering, University College London, London WC1H 0AH, UK.
*
Author to whom correspondence should be addressed.
Received: 14 September 2018 / Accepted: 21 September 2018 / Published: 25 September 2018
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Abstract

Raman spectroscopy is a novel tool used in the on-line monitoring and control of bioprocesses, offering both quantitative and qualitative determination of key process variables through spectroscopic analysis. However, the wide-spread application of Raman spectroscopy analysers to industrial fermentation processes has been hindered by problems related to the high background fluorescence signal associated with the analysis of biological samples. To address this issue, we investigated the influence of fluorescence on the spectra collected from two Raman spectroscopic devices with different wavelengths and detectors in the analysis of the critical process parameters (CPPs) and critical quality attributes (CQAs) of a fungal fermentation process. The spectra collected using a Raman analyser with the shorter wavelength (903 nm) and a charged coupled device detector (CCD) was corrupted by high fluorescence and was therefore unusable in the prediction of these CPPs and CQAs. In contrast, the spectra collected using a Raman analyser with the longer wavelength (993 nm) and an indium gallium arsenide (InGaAs) detector was only moderately affected by fluorescence and enabled the generation of accurate estimates of the fermentation’s critical variables. This novel work is the first direct comparison of two different Raman spectroscopy probes on the same process highlighting the significant detrimental effect caused by high fluorescence on spectra recorded throughout fermentation runs. Furthermore, this paper demonstrates the importance of correctly selecting both the incident wavelength and detector material type of the Raman spectroscopy devices to ensure corrupting fluorescence is minimised during bioprocess monitoring applications. View Full-Text
Keywords: Raman spectroscopy; fluorescence; fermentation monitoring; PLS modelling; critical process parameters; critical quality attributes Raman spectroscopy; fluorescence; fermentation monitoring; PLS modelling; critical process parameters; critical quality attributes
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Goldrick, S.; Lovett, D.; Montague, G.; Lennox, B. Influence of Incident Wavelength and Detector Material Selection on Fluorescence in the Application of Raman Spectroscopy to a Fungal Fermentation Process. Bioengineering 2018, 5, 79.

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