Comparison of colorimetric, fluorometric, and liquid chromatography-mass spectrometry assays for acetyl-coenzyme A.

Autor: Kantner DS; Lewis Katz School of Medicine at Temple University, Department of Cardiovascular Sciences, Aging + Cardiovascular Discovery Center, Philadelphia, PA, 19140, USA., Megill E; Lewis Katz School of Medicine at Temple University, Department of Cardiovascular Sciences, Aging + Cardiovascular Discovery Center, Philadelphia, PA, 19140, USA., Bostwick A; Lewis Katz School of Medicine at Temple University, Department of Cardiovascular Sciences, Aging + Cardiovascular Discovery Center, Philadelphia, PA, 19140, USA., Yang V; Lewis Katz School of Medicine at Temple University, Department of Cardiovascular Sciences, Aging + Cardiovascular Discovery Center, Philadelphia, PA, 19140, USA., Bekeova C; MitoCare Center, Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, Philadelphia, PA, 19107, USA., Van Scoyk A; Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, 84112, USA., Seifert EL; MitoCare Center, Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, Philadelphia, PA, 19107, USA., Deininger MW; Versiti Blood Research Institute and Medical College of Wisconsin, Milwaukee, WI, 53226, USA., Snyder NW; Lewis Katz School of Medicine at Temple University, Department of Cardiovascular Sciences, Aging + Cardiovascular Discovery Center, Philadelphia, PA, 19140, USA. Electronic address: NateWSnyder@temple.edu.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2024 Jan 15; Vol. 685, pp. 115405. Date of Electronic Publication: 2023 Nov 26.
DOI: 10.1016/j.ab.2023.115405
Abstrakt: Acetyl-Coenzyme A is a central metabolite in catabolic and anabolic pathways as well as the acyl donor for acetylation reactions. Multiple quantitative measurement techniques for acetyl-CoA have been reported, including commercially available kits. Comparisons between techniques for acetyl-CoA measurement have not been reported. This lack of comparability between assays makes context-specific assay selection and interpretation of results reporting changes in acetyl-CoA metabolism difficult. We compared commercially available colorimetric ELISA and fluorometric enzymatic-based kits to liquid chromatography-mass spectrometry-based assays using tandem mass spectrometry (LC-MS/MS) and high-resolution mass spectrometry (LC-HRMS). The colorimetric ELISA kit did not produce interpretable results even with commercially available pure standards. The fluorometric enzymatic kit produced comparable results to the LC-MS-based assays depending on matrix and extraction. LC-MS/MS and LC-HRMS assays produced well-aligned results, especially when incorporating stable isotope-labeled internal standards. In addition, we demonstrated the multiplexing capability of the LC-HRMS assay by measuring a suite of short-chain acyl-CoAs in a variety of acute myeloid leukemia cell lines and patient cells.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023. Published by Elsevier Inc.)
Databáze: MEDLINE
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