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 -