Integrated sampling procedure for metabolome analysis
Integrated sampling procedure for metabolome analysis
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DOI：
10.1021/bp050381q
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发表时间：
2006-10-06
影响因子：
2.9
通讯作者：
Dauner, Michael
中科院分区：
文献类型：
--
作者：
Schaub, Jochen;Schiesling, Carola;Dauner, Michael
代谢组分析是对大量细胞内代谢物的分析，已成为生物技术和药学研究中一种重要的系统分析方法。在代谢工程中，将代谢组数据与流动组和蛋白质组数据集成到大规模数学模型中，有望为菌株和细胞系的改进制定合理的策略。然而，开发用于定量分析细胞内代谢物浓度的可重复采样程序是一个重大挑战，实现(I)快速转移样品，(Ii)有效地猝灭新陈代谢，(Iii)定量提取代谢物，以及(Iv)为随后的定量分析优化样品条件。为了满足这些要求，我们提出了一个完整的抽样程序。通过将细胞短期暴露在温度下，实现了细胞内代谢物的同时猝灭和定量提取
Metabolome analysis, the analysis of large sets of intracellular metabolites, has become an important systems analysis method in biotechnological and pharmaceutical research. In metabolic engineering, the integration of metabolome data with fluxome and proteome data into large-scale mathematical models promises to foster rational strategies for strain and cell line improvement. However, the development of reproducible sampling procedures for quantitative analysis of intracellular metabolite concentrations represents a major challenge, accomplishing (i) fast transfer of sample, (ii) efficient quenching of metabolism, (iii) quantitative metabolite extraction, and (iv) optimum sample conditioning for subsequent quantitative analysis. In addressing these requirements, we propose an integrated sampling procedure. Simultaneous quenching and quantitative extraction of intracellular metabolites were realized by short-time exposure of cells to temperatures