Monitoring utilizations of amino acids and vitamins in culture media and Chinese hamster ovary cells by liquid chromatography tandem mass spectrometry

Qiu, Jinshu; Chan, Pik Kay; Bondarenko, Pavel V.

doi:10.1016/j.jpba.2015.08.036

Cited by 10 publications

(6 citation statements)

References 29 publications

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“…However, in their method Qiu et al . used an acidification sample preparation step to achieve sufficient peak shape with ion pairing chromatography on a reversed phase C18 column, which can potentially alter the chemical balance of the sample prior to its analysis . Floris et al .…”

Section: Introductionmentioning

confidence: 99%

“…However, in their method Qiu et al used an acidification sample preparation step to achieve sufficient peak shape with ion pairing chromatography on a reversed phase C18 column, which can potentially alter the chemical balance of the sample prior to its analysis. 32 Floris et al used nontarget MS to investigate the thermal stability of media during high-temperature short-term (HTST) sterilization, focusing particularly on Maillard products. 33 Among the multiple factors that are critical for CDM component stability, experience from 12 000-L scale manufacturing showed that preparation temperature is one of the main factors contributing to interbatch variability (unpublished data).…”

Section: Introductionmentioning

confidence: 99%

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Effect of manufacturing temperature and storage duration on stability of chemically defined media measured with LC‐MS/MS

Krattenmacher

Heermann

Calvet

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND Chemically defined media (CDM) are used routinely in industrial settings for the production of biopharmaceuticals from mammalian cells, so attention has shifted to understanding their chemical behavior and impact on process robustness. In this context, one particular problem is the risk of cross reactivity and instability in complex mixtures of chemicals at high concentrations. RESULTS In order to characterize potentially unstable medium compounds in feed media, we developed and validated a novel liquid chromatography tandem mass spectrometry (LC‐MS/MS) method to simultaneously quantify the majority of medium compounds. We used this application advantage to provide the following insights into molecular processes occurring in CDM prepared at different temperatures and during storage: Concentration profiles of critical compounds in CDM are independent of preparation temperature and storage material but show significant alterations over time. In particular l‐cysteine, l‐proline, vitamin B6, thiamine and cyanocobalamin were found to be rather unstable. Using mixed mode chromatography allows expansion of the method to cover additional analytes of interest, as demonstrated by inclusion of seven additional CDM reaction products. CONCLUSION The successful development of a new LC‐MS/MS method allowed us to show the impact of storage duration on the stability of CDM compounds. Furthermore, the results suggest that essential parameters for medium preparation (e.g. temperature) and storage upscaling (e.g. vessel material) are not impacting upon the chemical composition of CDM. © 2018 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effect of manufacturing temperature and storage duration on stability of chemically defined media measured with LC‐MS/MS

Krattenmacher

Heermann

Calvet

et al. 2018

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

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“…Due to the complexity of chemically defined medium, advanced technologies have become essential to comprehensively understand and further develop media. Liquid chromatography tandem mass spectrometry (LC–MS/MS) represents one such technology that is commonly used to quantify multiple AAs simultaneously 31 . Emerging technologies have also been recently utilized to identify and assess the quality of complex cell culture medium components, and these technologies include Raman spectroscopy, fluorescence excitation‐emission matrix spectroscopy (EEM), nuclear magnetic resonance spectroscopy (NMR), near‐infrared fluorescence spectroscopy (NIR), and thermogravimetric analysis (TGA) 32‐36 .…”

Section: Discussionmentioning

confidence: 99%

“…Liquid chromatography tandem mass spectrometry (LC-MS/MS) represents one such technology that is commonly used to quantify multiple AAs simultaneously. 31 Emerging technologies have also been recently utilized to identify and assess the quality of complex cell culture medium components, and these technologies include Raman spectroscopy, fluorescence excitation-emission matrix spectroscopy (EEM), nuclear magnetic resonance spectroscopy (NMR), near-infrared fluorescence spectroscopy (NIR), and thermogravimetric analysis (TGA). [32][33][34][35][36] As these technologies become readily available for manufacturing, they could be implemented in medium preparation to improve the process's robustness.…”

Section: Discussionmentioning

confidence: 99%

Impact of preparation pH and temperature on amino acid stability of highly concentrated cell culture feed media

Cui

et al. 2022

J of Chemical Tech & Biotech

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BACKGROUND The stability of amino acids (AAs) in a highly concentrated cell culture medium is affected by medium preparation and storage conditions. Variability in the concentration of medium components (including AAs) can influence cell culture performance and product quality attributes. RESULTS In this report, the impact of preparation pH, preparation temperature, and storage temperature was evaluated on the stability of 16 AAs in a highly concentrated cell culture feed medium. Optimizing the medium pH enabled an increase in the initial feed medium concentration by 30% plus 3.9 g L−1 tyrosine with no observed precipitation after 28 days. Up to 6.0 g L−1 tyrosine can be supplemented in the medium at ~pH 10 without precipitation. Moreover, preparing the medium at 40–50 °C prevented precipitation during storage at 4–8 °C. A slight negative impact on AA stability during this high‐temperature preparation was observed, but this impact was negligible relative to the AA degradation observed during extended storage of media prepared at 37 °C. The optimal preparation pH and temperature allowed the concentrated medium to be stored at 4–8 °C with minimal AA degradation and without precipitation. These discoveries were further supported by principal component analysis of AA profile similarity before and after storage at different pH and temperature conditions. CONCLUSION This study describes the medium preparation parameters to improve AA stability and mitigate precipitation issues. The optimization strategy will facilitate the development of robust cell culture production processes. © 2022 Society of Chemical Industry (SCI).

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“…Only very limited applications for vitamin analysis in biopharmaceutical samples are published. Some applications employ liquid chromatography coupled to mass spectrometry [13,14]. As these techniques are challenging to miniaturise, they are out of scope for the monitoring platform.…”

Section: Introductionmentioning

confidence: 99%

Design of experiments for micellar electrokinetic chromatography method development for the monitoring of water‐soluble vitamins in cell culture medium

van der Burg,

Wätzig,

Sänger – van de Griend

2023

Electrophoresis

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Biopharmaceutical production takes place in complex processes which should be thoroughly understood. Therefore, the iConsensus project focuses on developing a monitoring platform integrating several process analytical technology tools for integrated, automated monitoring of the biopharmaceutical process. Water‐soluble vitamin monitoring using (microchip) capillary electrophoresis (CE) is part of this platform. This work comprises the development of conventional CE methods as the first part towards integrated vitamin monitoring. The vitamins were divided based on their physical–chemical properties to develop two robust methods. Previously, a method for the analysis of cationic vitamins (pyridoxine, pyridoxal, pyridoxamine, thiamine and nicotinamide) in cell culture medium was developed. This work focused on the development of a micellar electrokinetic chromatography method for anionic and neutral vitamins (riboflavin, d‐calcium pantothenate, biotin, folic acid, cyanocobalamin and ascorbic acid). By employing multivariate design of experiments, the background electrolyte (BGE) could be optimised within one experiment testing only 11 BGEs. The optimised BGE conditions were 200 mM borate with 77 mM sodium dodecyl sulphate at a pH of 8.6. Using this BGE, all above‐mentioned cationic, anionic and neutral vitamins could be separated in clean samples. In cell culture medium, most anionic and neutral vitamins could be separated. Combining the two methods allows for analysis of cationic, anionic and neutral vitamins in cell culture medium samples. The next step towards integrated vitamin monitoring includes transfer to microchip CE. Due to the lack of fast and reliable methods for vitamin monitoring, the developed capillary methods could be valuable as stand‐alone at‐line process analytical technology solutions as well.

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Monitoring utilizations of amino acids and vitamins in culture media and Chinese hamster ovary cells by liquid chromatography tandem mass spectrometry

Cited by 10 publications

References 29 publications

Effect of manufacturing temperature and storage duration on stability of chemically defined media measured with LC‐MS/MS

Effect of manufacturing temperature and storage duration on stability of chemically defined media measured with LC‐MS/MS

Impact of preparation pH and temperature on amino acid stability of highly concentrated cell culture feed media

Design of experiments for micellar electrokinetic chromatography method development for the monitoring of water‐soluble vitamins in cell culture medium

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