Related ArticlesAn NMR-based Assay for the ex vivo Determination of Soluble CD73 Activity in Serum.
Anal Chem. 2020 Sep 28;:
Authors: He Y, Zhao Y, Nelson DM, Klippel A, Reily MD
Abstract
Extracellular adenosine, produced through the activity of the ecto-5'-nucleotidase CD73, elicits potent immune-suppressive effects and its upregulation in tumor cells as well as in stromal and immune cell subsets within the tumor microenvironment is hypothesized to represent an important resistance mechanism to current cancer immunotherapies. Soluble CD73 (sCD73) enzymatic activity measured in patient serum or plasma at baseline is reported to have prognostic as well as predictive relevance, with higher sCD73 activity associating with poor overall and progression-free survival in melanoma patients undergoing anti-PD1 monoclonal antibody treatment. Here we report a novel NMR-based method that measures the ex-vivo kinetics of sCD73 activity with high specificity and reproducibility and is suitable for future high-throughput implementation. Unlike existing assays, this method has the advantage of directly and simultaneously measuring the concentration of both, the CD73 substrate and product with minimal sample manipulation or special reagents. We establish the utility of the assay for measuring the activity of sCD73 in human serum and show a strong linear correlation between sCD73 protein levels and enzyme activity. Together with our finding that sCD73 appears to be the predominant activity for the generation of adenosine in human blood, our results demonstrate a link between activity and protein levels that will inform future clinical application.
PMID: 32985862 [PubMed - as supplied by publisher]
[NMR paper] Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay.
Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay.
Related Articles Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay.
J Clin Med. 2020 Sep 10;9(9):
Authors: Bedi S, Garcia E, Jeyarajah EJ, Shalaurova I, Perez-Matos MC, Jiang ZG, Dullaart RPF, Matyus SP, Kirk WJ, Otvos JD, Davidson WS, Connelly MA
Abstract
BACKGROUND: Lipoprotein particles with abnormal...
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09-17-2020 07:46 AM
[ASAP] Facile, Fluorogenic Assay for Protein Histidine Phosphatase Activity
Facile, Fluorogenic Assay for Protein Histidine Phosphatase Activity
https://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.8b00278/20180423/images/medium/bi-2018-00278c_0004.gif
Biochemistry
DOI: 10.1021/acs.biochem.8b00278
http://feeds.feedburner.com/~ff/acs/bichaw?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/bichaw/~4/XYxwyJU8rhA
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04-24-2018 01:29 PM
Fumarase activity: an in vivo and in vitro biomarker for acute kidney injury
From The DNP-NMR Blog:
Fumarase activity: an in vivo and in vitro biomarker for acute kidney injury
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Nielsen, P.M., et al., Fumarase activity: an in vivo and in vitro biomarker for acute kidney injury. Scientific Reports, 2017. 7: p. 40812.
http://dx.doi.org/10.1038/srep40812
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05-09-2017 12:37 AM
[NMR paper] Fluorine NMR-based assay in living mammalian cells.
Fluorine NMR-based assay in living mammalian cells.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Fluorine NMR-based assay in living mammalian cells.
Anal Biochem. 2015 Dec 11;
Authors: Veronesi M, Giacomina F, Romeo E, Castellani B, Ottonello G, Lambruschini C, Garau G, Scarpelli R, Bandiera T, Piomelli D, Dalvit C
Abstract
Nuclear Magnetic Resonance (NMR)-based screening has been recognized as a powerful approach for the identification...
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12-28-2015 12:26 AM
In vivo Overhauser-enhanced MRI of proteolytic activity
From The DNP-NMR Blog:
In vivo Overhauser-enhanced MRI of proteolytic activity
Koonjoo, N., et al., In vivo Overhauser-enhanced MRI of proteolytic activity. Contrast Media Mol Imaging, 2014. 9(5): p. 363-71.
http://www.ncbi.nlm.nih.gov/pubmed/24729587
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01-28-2015 05:28 PM
Cracking the molecular weight barrier: Fragment screening of an aminotransferase using an NMR-based functional assay.
Cracking the molecular weight barrier: Fragment screening of an aminotransferase using an NMR-based functional assay.
Cracking the molecular weight barrier: Fragment screening of an aminotransferase using an NMR-based functional assay.
Bioorg Med Chem Lett. 2011 Jul 21;
Authors: Mendoza R, Petros AM, Liu Y, Thimmapaya R, Surowy CS, Leise WF, Pereda-Lopez A, Panchal SC, Sun C
NMR-based screening of protein targets has become a well established part of the drug discovery process especially with respect to fragments. However, as target size increases...
A new method for the determination of free L: -carnitine in serum samples based on high field single quantum coherence filtering (1)H-NMR spectroscopy.
A new method for the determination of free L: -carnitine in serum samples based on high field single quantum coherence filtering (1)H-NMR spectroscopy.
A new method for the determination of free L: -carnitine in serum samples based on high field single quantum coherence filtering (1)H-NMR spectroscopy.
Anal Bioanal Chem. 2011 Jan 11;
Authors: Tsiafoulis CG, Exarchou V, Tziova PP, Bairaktari E, Gerothanassis IP, Troganis AN
The rapid and accurate determination of specific metabolites present in biofluids is a very demanding task which is essential...
An NMR-based Assay for the ex vivo Determination of Soluble CD73 Activity in Serum.
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