High-throughput nucleoside phosphate monitoring in mammalian cell fed-batch cultivation using quantitative matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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Author(s) Steinoff, Robert F., Ivarsson, Marija, Habicher, Tobias, Villiger, Thomas K., Boertz, Jens, Krismer, Jasmin, Fagerer, Stephan R., Soos, Miroslav, Morbidelli, Massimo, Pabst, Martin, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title High-throughput nucleoside phosphate monitoring in mammalian cell fed-batch cultivation using quantitative matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
Author(s) Steinoff, Robert F.
Ivarsson, Marija
Habicher, Tobias
Villiger, Thomas K.
Boertz, Jens
Krismer, Jasmin
Fagerer, Stephan R.
Soos, Miroslav
Morbidelli, Massimo
Pabst, Martin
Zenobi, Renato
Journal or Series Title Biotechnology Journal
Volume Number 10
Issue Number 1
Start Page 190
End Page 198
ISSN 1860-7314
Publisher Wiley-VCH
Publication Place Weinheim
Publication Date 2015-01
Keyword(s) ATP
Automated sample aliquoting
MALDI-MS
Microarrays for mass spectrometry (MAMS)
Monoclonal antibody production
Abstract Current methods for monitoring multiple intracellular metabolite levels in parallel are limited in sample throughput capabilities and analyte selectivity. This article presents a novel high-throughput method based on matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry (TOF-MS) for monitoring intracellular metabolite levels in fed-batch processes. The MALDI-TOF-MS method presented here is based on a new microarray sample target and allows the detection of nucleoside phosphates and various other metabolites using stable isotope labeled internal standards. With short sample preparation steps and thus high sample throughput capabilities, the method is suitable for monitoring mammalian cell cultures, such as antibody producing hybridoma cell lines in industrial environments. The method is capable of reducing the runtime of standard LC-UV methods to approximately 1 min per sample (including 10 technical replicates). Its performance is exemplarily demonstrated in an 8-day monitoring experiment of independently controlled fed-batches, containing an antibody producing mouse hybridoma cell culture. The monitoring profiles clearly confirmed differences between cultivation conditions. Hypothermia and hyperosmolarity were studied in four bioreactors, where hypothermia was found to have a positive effect on the longevity of the cell culture, whereas hyperosmolarity lead to an arrest of cell proliferation. The results are in good agreement with HPLC-UV cross validation experiments. Subsequent principal component analysis (PCA) clearly separates the different bioreactor conditions based on the measured mass spectral profiles. This method is not limited to any cell line and can be applied as a process analytical tool in biotechnological processes.
DOI 10.1002/biot.201400292
Additional Notes Received 1 May 2014, Revised 19 June 2014, Accepted 18 August 2014, Published online 18 September 2014
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
03451
Organisational Unit(s)
NEBIS System Number 010289881
Source Database ID FORM-1421324116
WOS-000348058200022
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@article{Stnff2015,
  author = "Steinoff, Robert F. and Ivarsson, Marija and Habicher, Tobias and Villiger, Thomas K. and Boertz, Jens and Krismer, Jasmin and Fagerer, Stephan R. and Soos, Miroslav and Morbidelli, Massimo and Pabst, Martin and Zenobi, Renato",
  title = "{H}igh-throughput nucleoside phosphate monitoring in mammalian cell fed-batch cultivation using quantitative matrix-assisted laser desorption/ionization time-of-flight mass spectrometry",
  journal = "Biotechnology Journal",
  year = 2015,
  volume = "10",
  number = "1",
  pages = "190--198",
  month = jan,
}


E-Citations record created: Thu, 15 Jan 2015, 12:15:23 CET