High-throughput LC-MS quantitation of cell culture metabolites

Sun, Zhonghua; Ji, Qinqin; Lewis, Michael; Mo, Jingjie; Hu, Ping

doi:10.1016/j.biologicals.2019.07.003

Cited by 8 publications

(3 citation statements)

References 58 publications

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“…This presents alanine as an ideal candidate as a low‐cost alternative to typical monoisotope IS. Another advantage of incorporating alanine as an IS is that the methyl side chain of alanine is widely known as inactive and, hence, is a stable compound in solution that generally can avoid the risk of being methylated into a different compound 14 . However, we appreciate that this method cannot be applied to samples from cell culture (since the cells may produce or consume the alanine changing the concentration) or media formulations that may contain alanine as an unknown.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Chinese hamster ovary cell culture feed media precipitate

Hoang

Galbraith

Kuang

et al. 2021

Biotechnology Progress

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Process intensification of monoclonal antibody production is leading to more concentrated feed media causing issues with precipitation of solids from the media solution. This results in processing problems since components in the precipitate are no longer in solution, changing the media composition and leading to variability in cell culture performance. The goal of this work is to characterize the feed media precipitate, and in particular to identify the precipitated components so that mitigation strategies can be developed. From the conducted analysis, the precipitate was predominately found to be organic and was analyzed with liquid chromatography‐mass spectrometry and inductively coupled plasma‐optical emission spectroscopy (ICP‐OES) to identify the constituent components. Up to ten amino acids were identified with tyrosine (approximately 77 wt.%) and phenylalanine (approximately 4 wt.%) being the most prevalent amino acids. Elemental analysis with ICP‐OES revealed that inorganic components were accounted for less than one weight percentage of the solid precipitate with metal sulfates being the predominant inorganic components.

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Section: Methodsmentioning

confidence: 99%

Characterization of Chinese hamster ovary cell culture feed media precipitate

Hoang

Galbraith

Kuang

et al. 2021

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…To circumvent these issues, uniform isotopic labeling techniques have been developed to aid in the confidence of metabolite identification in cells. 82 Isotope labeling techniques typically involve introducing 13 C-, 15 N-, or 18 O-labeled nutrients into the cell culture media. In MS-based metabolomics studies, 13 C-labeling techniques have been more commonly implemented, specifically for measuring metabolic flux of high-energy metabolites (e.g., glycolytic, TCA cycle, and pentose phosphate pathway intermediates).…”

Section: Sample Handling and Extraction Methodologymentioning

confidence: 99%

“…To date, the majority of MS-based cell research studies have analyzed cell population-averaged metabolic profiles, since most sampling mechanisms utilize large cell counts for sufficient MS sensitivity to reliably detect molecular alterations. To augment the information gained from analyzing intracellular metabolites, the exometabolome of cells can also be examined by sampling spent cell culture media, as it is an information-rich medium containing metabolites for monitoring cell-signaling processes . Population-averaged MS analyses, although apparently ubiquitous in the metabolomics literature, can be unrepresentative of individual cell-to-cell variability, masking unique cellular functions throughout a population of seemingly identical cells. , With modifications to existing instrumentation and the development of new sampling techniques, single-cell metabolome analyses are becoming more commonplace, revealing the complexity of cellular heterogeneity within subpopulations of cells .…”

mentioning

confidence: 99%