Microtechnology meets systems biology: The small molecules of metabolome as next big targets

Wurm, Matthias; Schöpke, Benedikt; Lutz, David; Müller, Jörg; Zeng, An‐Ping

doi:10.1016/j.jbiotec.2010.05.002

Cited by 22 publications

(15 citation statements)

References 156 publications

(257 reference statements)

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“…Shintu et al 355 made an organ-on-a-chip device that was probed via proton nuclear magnetic resonance for metabolomic analysis of liver and kidney cell cultures exposed to several toxic compounds. For further information on applications of microfluidic systems in metabolomics, the reader is referred to review articles by Wurm et al 356 and Kraly et al 357 …”

Section: Applicationsmentioning

confidence: 99%

Advances in Microfluidic Materials, Functions, Integration, and Applications

2013

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Section: Applicationsmentioning

confidence: 99%

Advances in Microfluidic Materials, Functions, Integration, and Applications

2013

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“…To investigate whether extended filtration times affect intracellular metabolite levels, whole cell filtrates were set aside for holding times of 0, 30 and 60 sec before freezing (after filtration) was carried out. Because ATP, ADP and AMP pool sizes are known to show fast turnover times [33,11], the adenylate pools were chosen to test whether metabolism is affected by different filtration durations (Fig. 1B).…”

Section: Effect Of Sample Time On Cellular Metabolites and Their Consmentioning

confidence: 99%

“…Experimental evidence that pool sizes between cytoplasm and mitochondrion may differ [9] have already been published. Being able to distinguish between cytosolic and mitochondrial pool sizes is not only important to describe cellular physiology but also to quantify intracellular flux distributions properly [10][11][12].…”

mentioning

confidence: 99%

Compartment‐specific metabolomics for CHO reveals that ATP pools in mitochondria are much lower than in cytosol

Matuszczyk

Teleki

Pfizenmaier

2015

Biotechnology Journal

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Mammalian cells show a compartmented metabolism. Getting access to subcellular metabolite pools is of high interest to understand the cells' metabolomic state. Therefore a protocol is developed and applied for monitoring compartment-specific metabolite and nucleotide pool sizes in Chinese hamster ovary (CHO) cells. The approach consists of a subtracting filtering method separating cytosolic components from physically intact mitochondrial compartments. The internal standards glucose-6-phosphate and cis-aconitate were chosen to quantify cytosolic secession and mitochondrial membrane integrity. Extracts of related fractions were studied by liquid chromatography-isotope dilution mass spectrometry for the absolute quantification of a subset of glycolytic and tricarboxylic acid cycle intermediates together with the adenylate nucleotides ATP, ADP and AMP. The application of the protocol revealed highly dynamic changes in the related pool sizes as a function of distinct cultivation periods of IgG1 producing CHO cells. Mitochondrial and cytosolic pool dynamics were in agreement with anticipated metabolite pools of independent studies. The analysis of adenosine phosphate levels unraveled significantly higher ATP levels in the cytosol leading to the hypothesis that mitochondria predominantly serve for fueling ATP into the cytosol where it is tightly controlled at physiological adenylate energy charges about 0.9.

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“…In addition, the absolute quantification of metabolites under physiological, in vivo and dynamic conditions remains a major challenge. The combination of existing multiparallel analytical platforms with special attention to metabolite quantification (see Sections 2.1.2 and 2.2.3) in a cohesive manner may not be sufficient and emerging microtechnologies such as microfluidics will certainly help (see Section 2.2.4 and (Wurm et al, 2010)). …”

Section: Systems Biologymentioning

confidence: 99%