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NMR methods for metabolomics of mammalian cell culture bioreactors. | LitMetric

NMR methods for metabolomics of mammalian cell culture bioreactors.

Methods Mol Biol

Bristol-Myers Squibb MS L14-05, Princeton, NJ, USA.

Published: August 2014

AI Article Synopsis

  • - Metabolomics utilizes NMR spectroscopy to analyze small molecules in mammalian cell cultures that produce therapeutic proteins, requiring little sample prep and no separation processes.
  • - NMR allows direct measurement of various small molecule concentrations in both the culture supernatant and inside the cells, which can be linked to important factors like protein yield, cell health, and glycosylation.
  • - The chapter emphasizes techniques for sample prep, data collection, and analysis specific to CHO cell cultures, with potential applications for similar in vitro systems.

Article Abstract

Metabolomics has become an important tool for measuring pools of small molecules in mammalian cell cultures expressing therapeutic proteins. NMR spectroscopy has played an important role, largely because it requires minimal sample preparation, does not require chromatographic separation, and is quantitative. The concentrations of large numbers of small molecules in the extracellular media or within the cells themselves can be measured directly on the culture supernatant and on the supernatant of the lysed cells, respectively, and correlated with endpoints such as titer, cell viability, or glycosylation patterns. The observed changes can be used to generate hypotheses by which these parameters can be optimized. This chapter focuses on the sample preparation, data acquisition, and analysis to get the most out of NMR metabolomics data from CHO cell cultures but could easily be extended to other in vitro culture systems.

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Source
http://dx.doi.org/10.1007/978-1-62703-733-4_15DOI Listing

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