title: Advanced online monitoring of cell culture off-gas using proton transfer reaction mass spectrometry.
abstract: Mass spectrometry has been frequently applied to monitor the O₂ and CO₂ content in the off-gas of animal cell culture fermentations. In contrast to classical mass spectrometry the proton transfer reaction mass spectrometry (PTR-MS) provides additional information of volatile organic compounds by application of a soft ionization technology. Hence, the spectra show less fragments and can more accurately assigned to particular compounds. In order to discriminate between compounds of non-metabolic and metabolic origin cell free experiments and fed-batch cultivations with a recombinant CHO cell line were conducted. As a result, in total eight volatiles showing high relevance to individual cultivation or cultivation conditions could be identified. Among the detected compounds methanethiol, with a mass-to-charge ratio of 49, qualifies as a key candidate in process monitoring due to its strong connectivity to lactate formation. Moreover, the versatile and complex data sets acquired by PTR MS provide a valuable resource for statistical modeling to predict non direct measurable parameters. Hence, partial least square regression was applied to the complete spectra of volatiles measured and important cell culture parameters such as viable cell density estimated (R²  = 0.86). As a whole, the results of this study clearly show that PTR-MS provides a powerful tool to improve bioprocess-monitoring for mammalian cell culture. Thus, specific volatiles emitted by cells and measured online by the PTR-MS and complex variables gained through statistical modeling will contribute to a deeper process understanding in the future and open promising perspectives to bioprocess control.
authors.affiliation: Sandoz GmbH, 6336, Langkampfen, Austria; Dept. of Biotechnology, University of Natural Resources and Life Sciences, 1180, Vienna, Austria.
authors.forename: Timo
authors.initials: T
authors.lastname: Schmidberger
authors.forename: Rene
authors.initials: R
authors.lastname: Gutmann
authors.forename: Karl
authors.initials: K
authors.lastname: Bayer
authors.forename: Jennifer
authors.initials: J
authors.lastname: Kronthaler
authors.forename: Robert
authors.initials: R
authors.lastname: Huber
mesh_terms: D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D018929:Cell Culture Techniques; D006224:Cricetinae; D003412:Cricetulus; D005285:Fermentation / Q000502:physiology*; D019344:Lactic Acid / Q000032:analysis / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D055549:Volatile Organic Compounds / Q000032:analysis* / Q000378:metabolism*
chemical_list: D055549:Volatile Organic Compounds; D019344:Lactic Acid
keywords: CHO cell culture; bioprocess monitoring; proton transfer reaction mass spectrometry (PTR-MS); volatile organic compound