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The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids

Nicotinamide adenine dinucleotide (NAD), a prominent member of the pyridine nucleotide family, plays a pivotal role in cell-oxidation protection, DNA repair, cell signalling and central metabolic pathways, such as beta oxidation, glycolysis and the citric acid cycle. In particular, extracellular NAD...

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Autores principales: Brunnbauer, Philipp, Leder, Annekatrin, Kamali, Can, Kamali, Kaan, Keshi, Eriselda, Splith, Katrin, Wabitsch, Simon, Haber, Philipp, Atanasov, Georgi, Feldbrügge, Linda, Sauer, Igor M., Pratschke, Johann, Schmelzle, Moritz, Krenzien, Felix
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208386/
https://www.ncbi.nlm.nih.gov/pubmed/30382125
http://dx.doi.org/10.1038/s41598-018-34350-6
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author Brunnbauer, Philipp
Leder, Annekatrin
Kamali, Can
Kamali, Kaan
Keshi, Eriselda
Splith, Katrin
Wabitsch, Simon
Haber, Philipp
Atanasov, Georgi
Feldbrügge, Linda
Sauer, Igor M.
Pratschke, Johann
Schmelzle, Moritz
Krenzien, Felix
author_facet Brunnbauer, Philipp
Leder, Annekatrin
Kamali, Can
Kamali, Kaan
Keshi, Eriselda
Splith, Katrin
Wabitsch, Simon
Haber, Philipp
Atanasov, Georgi
Feldbrügge, Linda
Sauer, Igor M.
Pratschke, Johann
Schmelzle, Moritz
Krenzien, Felix
author_sort Brunnbauer, Philipp
collection PubMed
description Nicotinamide adenine dinucleotide (NAD), a prominent member of the pyridine nucleotide family, plays a pivotal role in cell-oxidation protection, DNA repair, cell signalling and central metabolic pathways, such as beta oxidation, glycolysis and the citric acid cycle. In particular, extracellular NAD(+) has recently been demonstrated to moderate pathogenesis of multiple systemic diseases as well as aging. Herein we present an assaying method, that serves to quantify extracellular NAD(+) in human heparinised plasma and exhibits a sensitivity ranging from the low micromolar into the low nanomolar domain. The assay achieves the quantification of extracellular NAD(+) by means of a two-step enzymatic cycling reaction, based on alcohol dehydrogenase. An albumin modified revised simulated body fluid was employed as standard matrix in order to optimise enzymatic activity and enhance the linear behaviour and sensitivity of the method. In addition, we evaluated assay linearity, reproducibility and confirmed long-term storage stability of extracellular NAD(+) in frozen human heparinised plasma. In summary, our findings pose a novel standardised method suitable for high throughput screenings of extracellular NAD(+) levels in human heparinised plasma, paving the way for new clinical discovery studies.
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spelling pubmed-62083862018-11-01 The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids Brunnbauer, Philipp Leder, Annekatrin Kamali, Can Kamali, Kaan Keshi, Eriselda Splith, Katrin Wabitsch, Simon Haber, Philipp Atanasov, Georgi Feldbrügge, Linda Sauer, Igor M. Pratschke, Johann Schmelzle, Moritz Krenzien, Felix Sci Rep Article Nicotinamide adenine dinucleotide (NAD), a prominent member of the pyridine nucleotide family, plays a pivotal role in cell-oxidation protection, DNA repair, cell signalling and central metabolic pathways, such as beta oxidation, glycolysis and the citric acid cycle. In particular, extracellular NAD(+) has recently been demonstrated to moderate pathogenesis of multiple systemic diseases as well as aging. Herein we present an assaying method, that serves to quantify extracellular NAD(+) in human heparinised plasma and exhibits a sensitivity ranging from the low micromolar into the low nanomolar domain. The assay achieves the quantification of extracellular NAD(+) by means of a two-step enzymatic cycling reaction, based on alcohol dehydrogenase. An albumin modified revised simulated body fluid was employed as standard matrix in order to optimise enzymatic activity and enhance the linear behaviour and sensitivity of the method. In addition, we evaluated assay linearity, reproducibility and confirmed long-term storage stability of extracellular NAD(+) in frozen human heparinised plasma. In summary, our findings pose a novel standardised method suitable for high throughput screenings of extracellular NAD(+) levels in human heparinised plasma, paving the way for new clinical discovery studies. Nature Publishing Group UK 2018-10-31 /pmc/articles/PMC6208386/ /pubmed/30382125 http://dx.doi.org/10.1038/s41598-018-34350-6 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Brunnbauer, Philipp
Leder, Annekatrin
Kamali, Can
Kamali, Kaan
Keshi, Eriselda
Splith, Katrin
Wabitsch, Simon
Haber, Philipp
Atanasov, Georgi
Feldbrügge, Linda
Sauer, Igor M.
Pratschke, Johann
Schmelzle, Moritz
Krenzien, Felix
The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
title The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
title_full The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
title_fullStr The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
title_full_unstemmed The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
title_short The nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
title_sort nanomolar sensing of nicotinamide adenine dinucleotide in human plasma using a cycling assay in albumin modified simulated body fluids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208386/
https://www.ncbi.nlm.nih.gov/pubmed/30382125
http://dx.doi.org/10.1038/s41598-018-34350-6
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