Details
| Original language | English |
|---|---|
| Pages (from-to) | 352-362 |
| Number of pages | 11 |
| Journal | Engineering in life sciences |
| Volume | 19 |
| Issue number | 5 |
| Early online date | 14 Mar 2019 |
| Publication status | Published - 6 May 2019 |
Abstract
A new two-dimensional fluorescence sensor system was developed for in-line monitoring of mammalian cell cultures. Fluorescence spectroscopy allows for the detection and quantification of naturally occurring intra- and extracellular fluorophores in the cell broth. The fluorescence signals correlate to the cells’ current redox state and other relevant process parameters. Cell culture pretests with twelve different excitation wavelengths showed that only three wavelengths account for a vast majority of spectral variation. Accordingly, the newly developed device utilizes three high-power LEDs as excitation sources in combination with a back-thinned CCD-spectrometer for fluorescence detection. This setup was first tested in a lab design of experiments study with process relevant fluorophores proving its suitability for cell culture monitoring with LOD in the μg/L range. The sensor was then integrated into a CHO-K1 cell culture process. The acquired fluorescence spectra of several batches were evaluated using multivariate methods. The resulting batch evolution models were challenged in deviating and “golden batch” validation runs. These first tests showed that the new sensor can trace the cells’ metabolic state in a fast and reliable manner. Cellular distress is quickly detected as a deviation from the “golden batch”.
Keywords
- 2D-fluorescence spectroscopy, CHO cell cultivation, in-line bioprocess monitoring, metabolism monitoring, MVDA, PAT
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
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In: Engineering in life sciences, Vol. 19, No. 5, 06.05.2019, p. 352-362.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - A novel LED-based 2D-fluorescence spectroscopy system for in-line monitoring of Chinese hamster ovary cell cultivations – Part I
AU - Graf, Alexander
AU - Claßen, Jens
AU - Solle, Dörte
AU - Hitzmann, Bernd
AU - Rebner, Karsten
AU - Hoehse, Marek
N1 - Funding Information: This research was supported by the German Federal Ministry of Education and Research (Grant-No. 13N12703). Furthermore, we are thankful for the great support from art photonics (Germany) and J&M Analytik AG (Germany) with the development of the probe and sensor system.
PY - 2019/5/6
Y1 - 2019/5/6
N2 - A new two-dimensional fluorescence sensor system was developed for in-line monitoring of mammalian cell cultures. Fluorescence spectroscopy allows for the detection and quantification of naturally occurring intra- and extracellular fluorophores in the cell broth. The fluorescence signals correlate to the cells’ current redox state and other relevant process parameters. Cell culture pretests with twelve different excitation wavelengths showed that only three wavelengths account for a vast majority of spectral variation. Accordingly, the newly developed device utilizes three high-power LEDs as excitation sources in combination with a back-thinned CCD-spectrometer for fluorescence detection. This setup was first tested in a lab design of experiments study with process relevant fluorophores proving its suitability for cell culture monitoring with LOD in the μg/L range. The sensor was then integrated into a CHO-K1 cell culture process. The acquired fluorescence spectra of several batches were evaluated using multivariate methods. The resulting batch evolution models were challenged in deviating and “golden batch” validation runs. These first tests showed that the new sensor can trace the cells’ metabolic state in a fast and reliable manner. Cellular distress is quickly detected as a deviation from the “golden batch”.
AB - A new two-dimensional fluorescence sensor system was developed for in-line monitoring of mammalian cell cultures. Fluorescence spectroscopy allows for the detection and quantification of naturally occurring intra- and extracellular fluorophores in the cell broth. The fluorescence signals correlate to the cells’ current redox state and other relevant process parameters. Cell culture pretests with twelve different excitation wavelengths showed that only three wavelengths account for a vast majority of spectral variation. Accordingly, the newly developed device utilizes three high-power LEDs as excitation sources in combination with a back-thinned CCD-spectrometer for fluorescence detection. This setup was first tested in a lab design of experiments study with process relevant fluorophores proving its suitability for cell culture monitoring with LOD in the μg/L range. The sensor was then integrated into a CHO-K1 cell culture process. The acquired fluorescence spectra of several batches were evaluated using multivariate methods. The resulting batch evolution models were challenged in deviating and “golden batch” validation runs. These first tests showed that the new sensor can trace the cells’ metabolic state in a fast and reliable manner. Cellular distress is quickly detected as a deviation from the “golden batch”.
KW - 2D-fluorescence spectroscopy
KW - CHO cell cultivation
KW - in-line bioprocess monitoring
KW - metabolism monitoring
KW - MVDA
KW - PAT
UR - http://www.scopus.com/inward/record.url?scp=85064015818&partnerID=8YFLogxK
U2 - 10.1002/elsc.201800149
DO - 10.1002/elsc.201800149
M3 - Article
AN - SCOPUS:85064015818
VL - 19
SP - 352
EP - 362
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 5
ER -