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Dissolution Dynamic Nuclear Polarization Instrumentation for Real-time Enzymatic Reaction Rate Measurements by NMR

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Published on: February 23, 2016

Stable Isotope-Resolved Analysis with Quantitative Dissolution Dynamic Nuclear Polarization.

Mathilde H Lerche¹, Demet Yigit¹, Anne B Frahm¹

¹Center for Hyperpolarization in Magnetic Resonance, Department of Electrical Engineering, Technical University of Denmark , Ørsteds Plads, 2800 Kongens Lyngby, Denmark.

Analytical Chemistry

|December 5, 2017

Summary

This summary is machine-generated.

This study introduces a method for quantitative dDNP NMR to analyze metabolic pathways using stable isotopes. This technique provides new insights into aggressive cancer cell metabolism.

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Area of Science:

Metabolomics
Nuclear Magnetic Resonance (NMR) Spectroscopy
Cancer Biology

Background:

NMR spectroscopy enables noninvasive analysis of metabolite profiles and isotopomer distributions in complex mixtures.
Dynamic Nuclear Polarization (dDNP) and isotope enrichment significantly enhance NMR sensitivity and resolution for metabolic studies.
Quantifying metabolic pathways and networks requires controlled protocols for ex situ signal enhancement.

Purpose of the Study:

To develop a reproducible and quantitative method for stable isotope-resolved analysis using dissolution Dynamic Nuclear Polarization (dDNP) NMR.
To apply this quantitative dDNP (qdDNP) NMR technique for investigating the metabolic phenotype of aggressive cancer cells.

Main Methods:

A comprehensive sample preparation protocol was established, including cell incubation, extraction, and signal enhancement.
Dissolution Dynamic Nuclear Polarization (dDNP) was employed to boost NMR signal sensitivity.
Stable isotope-resolved analysis was performed using quantitative dDNP (qdDNP) NMR.

Main Results:

A reproducible method for quantitative dDNP NMR-based stable isotope-resolved analysis was successfully developed.
The qdDNP NMR approach enabled the acquisition of detailed metabolic insights.
Metabolic characteristics of aggressive cancer cells were elucidated using the qdDNP NMR technique.

Conclusions:

The presented sample preparation and qdDNP NMR protocol allow for reproducible and quantitative metabolic analysis.
This technique offers a powerful tool for mapping and quantifying metabolic pathways and networks.
qdDNP NMR provides valuable metabolic insights into aggressive cancer cell phenotypes, advancing cancer research.