LABELED TRANSITION METAL COMPLEXES

• US 20100155590A1
• Filed: 12/01/2005
• Published: 06/24/2010
• Est. Priority Date: 12/08/2004
• Status: Active Grant

• Current Assignee: Leica Biosystems Newcastle Ltd. (Danaher Corporation)
• Original Assignee: Kreatech Biotechnology BV (Danaher Corporation)
• Inventors: Talman, Eduard Gerhard, Lacombe, Marie, Opdam, Franciscus Johannes, Veuskens, Jacky Theo Maria
• Granted Patent: US 9,188,590 B2
• Time in Patent Office: Days
• US Class Current: 250/282
• CPC Class Codes: C07F 15/0093 without a metal-carbon linkage
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence

Abstract

The invention relates to a labeled transition metal complex comprising a transition metal atom, a reactive moiety for allowing a chemical or biological entity to become attached to the transition metal atom, an inert tridentate moiety as a stabilizing bridge, and a marker. The invention also relates to a labeled chemical or biological entity comprising a chemical or biological entity which is attached to said labeled transition metal complex, to the use of said complex for creating a defined shift in the molecular mass of said entity in order to facilitate mass spectrometric analysis of said entity, to methods for rendering chemical or biological entities distinguishable by mass spectrometry as well as to methods for mass spectrometric analysis of the chemical or biological entities. In addition, the present invention also relates to a set of at least two of said transition metal complexes of different molecular mass, to transition metal complexes comprising different stable isotopes, to chemical or biological entities obtained by a method of the invention and to a kit of parts supporting the use and/or methods of the invention.

Claims

1. A labeled transition metal complex comprising a transition metal atom, a reactive moiety for allowing a chemical or biological entity to become attached to the transition metal atom, an inert tridentate moiety as a stabilizing bridge, and a marker.

2. A complex according to claim 1, wherein the transition metal is chosen from the group consisting of vanadium, chromium, iron, nickel, copper, ruthenium, palladium, platinum, molybdenum, tungsten, cobalt, manganese, osmium, rhodium, iridium, zinc, and cadmium.

3. A complex according to claim 2, wherein the transition metal is chosen from the group consisting of iron, nickel, ruthenium, palladium, platinum, molybdenum, tungsten, and cobalt.

4. A complex according to claim 3, wherein the transition metal is platinum, cobalt, or ruthenium.

5. A complex according to any one of claim 1, wherein the marker is contained in the tridentate moiety or attached to the tridentate moiety, optionally by means of a spacer.

6. A complex according to any one of claim 1, wherein the transition metal atom is attached to the tridentate moiety by means of three donor atoms, each being independently selected from the group of nitrogen, oxygen, sulphur, and phosphorus atoms, which donor atoms are present in the tridentate moiety.

7. A complex according to claim 6, wherein the transition metal atom is attached to the tridentate moiety by means of three nitrogen atoms.

8. A complex according to any one of claim 6 or 7, wherein the donor atoms in the tridentate moiety are separated from each other by 1 to 5, preferably 1 to 3 atoms.

9. A complex according to claim 1, wherein the marker is a fluorescent label.

10. A complex according to claim 1, wherein the reactive moiety is chosen from the group consisting of Cl⁻

, NO₃⁻

, HCO₃⁻

, CO₃²⁻

, SO₃²⁻

, ZSO₃⁻

, I⁻

, Br⁻

, F⁻

, acetate, carboxylate, phosphate, ethylnitrate, oxalate, citrate, a phosphonate, ZO⁻

, and water wherein z represents a hydrogen moiety or an alkyl or aryl group haying from 1 to 10 carbon atoms.

11. A labeled chemical or biological entity comprising a chemical or biological entity which is attached to a transition metal complex as defined in claim 1, wherein the chemical or biological entity has replaced the reactive moiety.

12. A labeled chemical or biological entity according to claim 11, wherein the entity is selected from the group consisting of amino acids, peptides, oligopeptide, polypeptides, proteins, immunoglobulins, enzymes, synzymes, phospholipides, glycoproteins, nucleic acids, nucleosides, nucleotides, oligonucleotides, polynucleotides, peptide nucleic acids, peptide nucleic acid oligomers, peptide nucleic acid polymers, amines and aminoglycosides.

13. A method for preparing a labeled chemical or biological entity according to claim 11 or 12, wherein a chemical or biological entity is brought in contact with a transition metal complex as defined in any one of claims 1-10 under such conditions that the reactive moiety is replaced by the chemical or biological entity.

14. Use of a transition metal complex as defined in claim 1, for creating a defined shift in the molecular weight of the chemical or biological entity in order to facilitate mass spectrometric analysis of said entity or a sample comprising said entity.

15. A method for rendering a chemical or biological entity distinguishable by mass spectrometry, said method comprising the step of differentially labeling said entity with at least one transition metal complex as defined in claim 1.

16. A method for mass spectrometric analysis of a chemical or biological entity, said method comprising the steps of differentially labeling said entity with at least one transition metal complex as defined in claim 1 and analyzing said entity by mass spectrometry.

17. A method according to claim 15 or 16, wherein at least two entities are labeled and wherein said entities originate from different samples.

18. A method according to claim 17, wherein said step of differentially labeling said entity is performed with at least two transition metal complexes as defined in any one of claims 1-9, wherein said transition metal complexes are of different molecular weight, said difference in molecular weight being ≧

1 Da.

19. A set of at least two transition metal complexes of different molecular weight, wherein each of said transition metal complexes is as defined in claim 1.

20. A kit of parts comprising the set of at least two transition metal complexes of different molecular weight as defined in claim 19.

21. Kit of parts according to claim 20, further comprising at least one item selected from the group consisting of reaction instructions, test samples, test tubes or strips, buffers, marker preparations and preparations for adjusting the ionic strength.

22. A kit of parts for employing a method according to any of the claims 15-18.