Evaluation of amino acid profile by targeted metabolomics in the eukaryotic model under exposure of benzo[a]pyrene as the exclusive stressor

Batista, Josimar Marques; Neves, Maria J.; Menezes, Helvécio Costa; Cardeal, Zenilda de Lourdes

doi:10.1016/j.talanta.2023.124859

Cited by 1 publication

(1 citation statement)

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“…For example, the extraction of metabolites using 100% methanol extraction followed by water was found to be most effective for the recovery of the largest range of metabolites in CHO cells by GC-MS ( Sellick et al, 2010 ). Derivatisation with propyl chloroformate/propanol obtains excellent extraction of amino acid analytes ( Batista et al, 2023 ). The recovery of fatty acids (e.g., stearic acid and palmitic acid) was maximized by hot ethanol extraction, while the recovery of glycerol-1-phosphate was significantly greater when cellular metabolites were extracted with KOH ( Park et al, 2021 ).…”

Section: Gaps Of Metabolomic Pipelinementioning

confidence: 99%

Advancements in CHO metabolomics: techniques, current state and evolving methodologies

Singh,

Fatima,

Thakur

et al. 2024

Front. Bioeng. Biotechnol.

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Background: Investigating the metabolic behaviour of different cellular phenotypes, i.e., good/bad grower and/or producer, in production culture is important to identify the key metabolite(s)/pathway(s) that regulate cell growth and/or recombinant protein production to improve the overall yield. Currently, LC-MS, GC-MS and NMR are the most used and advanced technologies for investigating the metabolome. Although contributed significantly in the domain, each technique has its own biasness towards specific metabolites or class of metabolites due to various reasons including variability in the concept of working, sample preparation, metabolite-extraction methods, metabolite identification tools, and databases. As a result, the application of appropriate analytical technique(s) is very critical.Purpose and scope: This review provides a state-of-the-art technological insights and overview of metabolic mechanisms involved in regulation of cell growth and/or recombinant protein production for improving yield from CHO cultures.Summary and conclusion: In this review, the advancements in CHO metabolomics over the last 10 years are traced based on a bibliometric analysis of previous publications and discussed. With the technical advancement in the domain of LC-MS, GC-MS and NMR, metabolites of glycolytic and nucleotide biosynthesis pathway (glucose, fructose, pyruvate and phenylalanine, threonine, tryptophan, arginine, valine, asparagine, and serine, etc.) were observed to be upregulated in exponential-phase thereby potentially associated with cell growth regulation, whereas metabolites/intermediates of TCA, oxidative phosphorylation (aspartate, glutamate, succinate, malate, fumarate and citrate), intracellular NAD+/NADH ratio, and glutathione metabolic pathways were observed to be upregulated in stationary-phase and hence potentially associated with increased cell-specific productivity in CHO bioprocess. Moreover, each of technique has its own bias towards metabolite identification, indicating their complementarity, along with a number of critical gaps in the CHO metabolomics pipeline and hence first time discussed here to identify their potential remedies. This knowledge may help in future study designs to improve the metabolomic coverage facilitating identification of the metabolites/pathways which might get missed otherwise and explore the full potential of metabolomics for improving the CHO bioprocess performances.

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Section: Gaps Of Metabolomic Pipelinementioning

confidence: 99%