Micromolar Concentration Affinity Study on a Benchtop NMR Spectrometer with Secondary ¹³C Labeled Hyperpolarized...

Area of Science:

• Biophysical Chemistry
• Analytical Chemistry
• Drug Discovery

Background:

• Benchtop NMR offers a low-cost alternative to high-field NMR but suffers from limited sensitivity.
• Hyperpolarization techniques, particularly dissolution dynamic nuclear polarization (dDNP), dramatically enhance NMR sensitivity.
• This sensitivity enhancement is crucial for applications like affinity studies in drug discovery.

Purpose of the Study:

• To demonstrate the use of 13C-labeled ligands with hyperpolarization for benchtop NMR.
• To assess the feasibility of using these hyperpolarized ligands for probing protein interactions.
• To evaluate the potential for accelerating drug candidate discovery.

Main Methods:

• Ligands were secondarily labeled with 13C tags.
• Hyperpolarization was achieved using conventional dissolution dynamic nuclear polarization (dDNP) methods.
• The hyperpolarized 13C-labeled ligands were analyzed using an 80 MHz benchtop NMR spectrometer.

Main Results:

• Hyperpolarization increased 13C NMR sensitivity by over 5 orders of magnitude.
• Hyperpolarized ligands exhibited long nuclear spin-lattice relaxation time constants.
• Detection of ligands at micromolar concentrations was achieved, simplifying spectra to single peaks per ligand.

Conclusions:

• Hyperpolarized 13C-labeled ligands enable sensitive detection on benchtop NMR systems.
• This approach facilitates probing molecular interactions relevant to drug discovery.
• The method holds promise for improving and accelerating the identification of new drug candidates.