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.

中文翻译：

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CHO 代谢组学的进展：技术、现状和不断发展的方法

背景：研究生产培养中不同细胞表型（即好/坏种植者和/或生产者）的代谢行为对于识别调节细胞生长和/或重组蛋白生产的关键代谢物/途径非常重要以提高整体产量。目前，LC-MS、GC-MS 和 NMR 是研究代谢组最常用和最先进的技术。尽管在该领域做出了重大贡献，但由于各种原因，包括工作概念、样品制备、代谢物提取方法、代谢物识别工具和数据库的可变性，每种技术对特定代谢物或代谢物类别都有自己的偏见。因此，应用适当的分析技术非常关键。 目的和范围：本综述提供了最先进的技术见解和对细胞生长和/或重组蛋白调节所涉及的代谢机制的概述摘要和结论：在这篇综述中，基于对先前出版物的文献计量分析，追溯了过去 10 年 CHO 代谢组学的进展并进行了讨论。随着LC-MS、GC-MS和NMR领域的技术进步，糖酵解和核苷酸生物合成途径的代谢物（葡萄糖、果糖、丙酮酸和苯丙氨酸、苏氨酸、色氨酸、精氨酸、缬氨酸、天冬酰胺和丝氨酸等）越来越多。观察到在指数期上调，因此可能与细胞生长调节相关，而 TCA 的代谢物/中间体、氧化磷酸化（天冬氨酸、谷氨酸、琥珀酸、苹果酸、富马酸和柠檬酸）、细胞内 NAD+/NADH 比率和谷胱甘肽代谢途径观察到在稳定期上调，因此可能与 CHO 生物过程中细胞特异性生产力的增加有关。此外，每种技术对代谢物鉴定都有自己的偏见，表明它们的互补性，以及 CHO 代谢组学管道中的许多关键差距，因此在这里首次讨论以确定其潜在的补救措施。这些知识可能有助于未来的研究设计，以提高代谢组学覆盖率，从而促进识别可能会被错过的代谢物/途径，并探索代谢组学在改善 CHO 生物过程性能方面的全部潜力。