NMR-based methods for detecting ligands, where the ligand or target are hyperpolarized and the NMR-spectrum is compared with a reference spectrum of the ligand or target
View Patent ↗The invention relates to NMR-based methods comprising hyperpolarised ligands or targets for use in NMR-based assays.
1. An NMR-based method comprising the steps of:
a) hyperpolarising at least one of a ligand and a target, wherein said at least one of a ligand and a target is isotopically enriched with at least one of 12 C and 15 N NMR active nuclei,
b) forming a mixture by contacting either the at least one hyperpolarised ligand with one of a target and both a target and at least one further ligand or the hyperpolarised target with at least one ligand,
c) generating a NMR spectrum of the mixture, and
d) comparing said NMR spectrum with a reference spectrum of the at least one hyperpolarised ligand or the hyperpolarised target.
2. The method according to claim 1 , wherein the at least one ligand is selected from the group consisting of proteins, glycoproteins, lipoproteins, polypeptides, glyco-polypeptides, lipopolypeptides, peptides, carbohydrates, nucleic acids or a part, a fragment or a complex thereof and small organic molecules.
3. The method according to claim 1 , wherein the at least one ligand is a small organic molecule of less than 2000 Da.
4. The method according to claim 1 , wherein more than one hyperpolarised ligand is used.
5. The method according to claim 1 , wherein the target is selected from the group consisting of proteins, glycoproteins, lipoproteins, nucleic acids, polypeptides, glycopolypeptides, lipopolypeptides, peptides or a part, a fragment or a complex thereof.
6. The method according to claim 1 , wherein the enrichment is a hu 13 C-enrichment.
7. The method according to claim 1 , wherein the NMR spectrum generated is a one-dimensional NMR spectrum.
8. The method according to claim 1 , wherein the NMR spectrum generated is generated using low flip angles.
9. The method according to claim 1 , wherein the comparison with the reference spectrum shows a chemical shift difference, a relaxation time difference or a NOE effect difference.