Related ArticlesNMR methods for metabolomics of Mammalian cell culture bioreactors.
Methods Mol Biol. 2014;1104:223-36
Authors: Aranibar N, Reily MD
Abstract
Metabolomics has become an important tool for measuring pools of small molecules in mammalian cell cultures expressing therapeutic proteins. NMR spectroscopy has played an important role, largely because it requires minimal sample preparation, does not require chromatographic separation, and is quantitative. The concentrations of large numbers of small molecules in the extracellular media or within the cells themselves can be measured directly on the culture supernatant and on the supernatant of the lysed cells, respectively, and correlated with endpoints such as titer, cell viability, or glycosylation patterns. The observed changes can be used to generate hypotheses by which these parameters can be optimized. This chapter focuses on the sample preparation, data acquisition, and analysis to get the most out of NMR metabolomics data from CHO cell cultures but could easily be extended to other in vitro culture systems.
[NMR paper] Site-specific NMR mapping and time-resolved monitoring of serine and threonine phosphorylation in reconstituted kinase reactions and mammalian cell extracts.
Site-specific NMR mapping and time-resolved monitoring of serine and threonine phosphorylation in reconstituted kinase reactions and mammalian cell extracts.
Site-specific NMR mapping and time-resolved monitoring of serine and threonine phosphorylation in reconstituted kinase reactions and mammalian cell extracts.
Nat Protoc. 2013 Jun 27;8(7):1416-1432
Authors: Theillet FX, Rose HM, Liokatis S, Binolfi A, Thongwichian R, Stuiver M, Selenko P
Abstract
We outline NMR protocols for site-specific mapping and time-resolved monitoring of...
NMR-based metabolomics of mammalian cell and tissue cultures
NMR-based metabolomics of mammalian cell and tissue cultures
Abstract NMR spectroscopy was used to evaluate growth media and the cellular metabolome in two systems of interest to biomedical research. The first of these was a Chinese hamster ovary cell line engineered to express a recombinant protein. Here, NMR spectroscopy and a quantum mechanical total line shape analysis were utilized to quantify 30 metabolites such as amino acids, Krebs cycle intermediates, activated sugars, cofactors, and others in both media and cell extracts. The impact of bioreactor scale and addition of...
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NMR-based metabolomics of mammalian cell and tissue cultures.
NMR-based metabolomics of mammalian cell and tissue cultures.
NMR-based metabolomics of mammalian cell and tissue cultures.
J Biomol NMR. 2011 Mar 4;
Authors: Aranibar N, Borys M, Mackin NA, Ly V, Abu-Absi N, Abu-Absi S, Niemitz M, Schilling B, Li ZJ, Brock B, Russell RJ, Tymiak A, Reily MD
NMR spectroscopy was used to evaluate growth media and the cellular metabolome in two systems of interest to biomedical research. The first of these was a Chinese hamster ovary cell line engineered to express a recombinant protein. Here, NMR spectroscopy and a...
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03-05-2011 01:02 PM
[NMR paper] NMR studies of the most conserved RNA domain of the mammalian signal recognition part
NMR studies of the most conserved RNA domain of the mammalian signal recognition particle (SRP).
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles NMR studies of the most conserved RNA domain of the mammalian signal recognition particle (SRP).
RNA. 1996 Dec;2(12):1213-27
Authors: Schmitz U, Freymann DM, James TL, Keenan RJ, Vinayak R, Walter P
Mammalian signal recognition particle (SRP) and its homologues exhibit a phylogenetically conserved RNA domain, whose...
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[NMR paper] Two-dimensional, rotational-echo double-resonance NMR of cell culture metabolism.
Two-dimensional, rotational-echo double-resonance NMR of cell culture metabolism.
Related Articles Two-dimensional, rotational-echo double-resonance NMR of cell culture metabolism.
J Biol Chem. 1993 Oct 5;268(28):20768-71
Authors: McDowell LM, Cohen ER, Schaefer J
Two-dimensional, rotational-echo double-resonance 13C NMR, a new solid-state NMR technique, has been used to show that the relative fluxes of the labeled chemical bond of L-serine along four metabolic pathways (direct purine synthesis, direct glycine incorporation into protein,...
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[NMR paper] Detection of fructose-3-phosphokinase activity in intact mammalian lenses by 31P NMR
Detection of fructose-3-phosphokinase activity in intact mammalian lenses by 31P NMR spectroscopy.
Related Articles Detection of fructose-3-phosphokinase activity in intact mammalian lenses by 31P NMR spectroscopy.
J Biol Chem. 1993 Apr 15;268(11):7763-7
Authors: Lal S, Szwergold BS, Kappler F, Brown T
Recently we have identified two novel phosphorylated metabolites in the lenses of diabetic rats as sorbitol 3-phosphate (Sor-3-P) and fructose 3-phosphate (Fru-3-P). The latter compound is of particular interest since it is a potent glycating...
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[NMR paper] Cadmium-113 NMR studies of the DNA binding domain of the mammalian glucocorticoid rec
Cadmium-113 NMR studies of the DNA binding domain of the mammalian glucocorticoid receptor.
Related Articles Cadmium-113 NMR studies of the DNA binding domain of the mammalian glucocorticoid receptor.
Biochemistry. 1990 Oct 2;29(39):9218-25
Authors: Pan T, Freedman LP, Coleman JE
The DNA binding domain of the mammalian glucocorticoid hormone receptor (GR) contains nine highly conserved cysteine residues, a conservation shared by the superfamily of steroid and thyroid hormone receptors. A fragment consisting of GR residues 407-556, containing...
NMR methods for metabolomics of Mammalian cell culture bioreactors.
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