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Direct optical detection of cell density and viability of mammalian cells by means of UV/VIS spectroscopy

  • The critical process parameters cell density and viability during mammalian cell cultivation are assessed by UV/VIS spectroscopy in combination with multivariate data analytical methods. This direct optical detection technique uses a commercial optical probe to acquire spectra in a label-free way without signal enhancement. For the cultivation, an inverse cultivation protocol is applied, which simulates the exponential growth phase by exponentially replacing cells and metabolites of a growing Chinese hamster ovary cell batch with fresh medium. For the simulation of the death phase, a batch of growing cells is progressively replaced by a batch with completely starved cells. Thus, the most important parts of an industrial batch cultivation are easily imitated. The cell viability was determined by the well-established method partial least squares regression (PLS). To further improve process knowledge, the viability has been determined from the spectra based on a multivariate curve resolution (MCR) model. With this approach, the progress of the cultivations can be continuously monitored solely based on an UV/VIS sensor. Thus, the monitoring of critical process parameters is possible inline within a mammalian cell cultivation process, especially the viable cell density. In addition, the beginning of cell death can be detected by this method which allows us to determine the cell viability with acceptable error. The combination of inline UV/VIS spectroscopy with multivariate curve resolution generates additional process knowledge complementary to PLS and is considered a suitable process analytical tool for monitoring industrial cultivation processes.

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Metadaten
Name:Drieschner, Tobias; Ostertag, Edwin; Boldrini, Barbara; Lorenz, Anita; Brecht, Marc; Rebner, Karsten
DOI:https://doi.org/10.1007/s00216-019-02322-y
ISSN:1618-2642
eISSN:1618-2650
Erschienen in:Analytical and bioanalytical chemistry : a merger of Fresenius' journal of analytical chemistry and Analusis
Publisher:Springer
Place of publication:Berlin ; Heidelberg
Document Type:Article
Language:English
Year of Publication:2020
Tag:UV/VIS spectroscopy; cell viability; multivariate curve resolution; partial least squares regression; process analytical technology
Volume:412
Issue:14
Pagenumber:13
First Page:3359
Last Page:3371
Dewey Decimal Classification:540 Chemie
Open Access:Nein
Licence (German):License Logo  Lizenzbedingungen Springer