A novel LED-based 2D-fluorescence spectroscopy system for in-line monitoring of Chinese hamster ovary cell cultivations - Part I.
ENGINEERING IN LIFE SCIENCES(2019)
摘要
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 g/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.
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关键词
2D-fluorescence spectroscopy,CHO cell cultivation,in-line bioprocess monitoring,metabolism monitoring,MVDA,PAT
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