IMMUNOASSAYS EMPLOYING NON-PARTICULATE CHEMILUMINESCENT REAGENT

• US 20100233725A1
• Filed: 03/12/2009
• Published: 09/16/2010
• Est. Priority Date: 03/12/2009
• Status: Active Grant

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Inventors: Geng, Liping, Janzen, Roland, Schelp, Carsten, Singh, Pratap, Zheng, Yi Feng
• Granted Patent: US 9,347,947 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: G01N 2333/59 Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/78 Thyroid gland hormones , e.g. T3, T4, TBH, TBG or their receptors

Abstract

Methods and reagents are disclosed for conducting assays. Embodiments of the present methods and reagents are concerned with chemiluminescent reagents for determining the presence and/or amount of an analyte in a sample suspected of containing the analyte. The reagent is non-particulate and comprises a binding partner for the analyte and a chemiluminescent composition comprising an olefinic compound and a metal chelate. In embodiments of an assay, a combination is provided that comprises a sample suspected of containing the analyte, a chemiluminescent reagent as described above and a sensitizer reagent capable of generating singlet oxygen. The combination is subjected to conditions for binding of the analyte to the binding partner for the analyte. The sensitizer is activated and the amount of luminescence generated by the chemiluminescent composition is detected wherein the amount of luminescence is related to the amount of the analyte in the sample.

Claims

1. A chemiluminescent reagent for determining the presence and/or amount of an analyte in a sample suspected of containing the analyte, the reagent being non-particulate and comprising a binding partner for the analyte and a chemiluminescent composition comprising an olefinic compound and a metal chelate.

2. The chemiluminescent reagent according to claim 1 wherein the binding partner for the analyte is covalently bound to the chemiluminescent composition by a bond or a linking group.

3. The chemiluminescent reagent according to claim 2 wherein the binding partner for the analyte is covalently bound to the chemiluminescent composition by a linking group and wherein the linking group comprises a hydrophilic macromolecule.

4. The chemiluminescent reagent according to claim 3 wherein the olefinic compound and the metal chelate are linked to the same molecule of the linking group to which the binding partner for the analyte is covalently bound.

5. The chemiluminescent reagent according to claim 3 wherein the olefinic compound and the metal chelate are linked to separate molecules of the linking group to which the binding partner for the analyte is covalently bound and wherein the linking group may be the same or different.

6. The chemiluminescent reagent according to claim 3 wherein the hydrophilic macromolecule is selected from the group consisting of polypeptides, dendrimers, polycarboxylates, polyamines, polysulfhydryls and polyethyleneglycols.

7. The chemiluminescent reagent according to claim 1 wherein the binding partner for the analyte is non-covalently bound to the chemiluminescent composition by a specific binding pair wherein one member of the specific binding pair is linked to the binding partner for the analyte and the other member of the specific binding pair is linked to the chemiluminescent composition by a bond or a linking group.

8. The chemiluminescent reagent according to claim 7 wherein the binding partner for the analyte is non-covalently bound to the chemiluminescent composition by a specific binding pair wherein one member of the specific binding pair is linked to the binding partner for the analyte and the other member of the specific binding pair is linked to the chemiluminescent composition by a linking group wherein the linking group is a protein.

9. The chemiluminescent reagent according to claim 8 wherein the olefinic compound and the metal chelate are linked to the same molecule of the linking group to which the binding partner for the analyte is non-covalently bound.

10. The chemiluminescent reagent according to claim 8 wherein the olefinic compound and the metal chelate are linked to separate molecules of the linking group to which the binding partner for the analyte is non-covalently bound and wherein the linking group may be the same or different.

11. The chemiluminescent reagent according to claim 1 wherein the hydrophilic macromolecule is a protein.

12. The chemiluminescent reagent according to claim 7 wherein the specific binding pair is selected from the group consisting of (i) small molecule and binding partner for the small molecule and (ii) large molecule and binding partner for the large molecule.

13. The chemiluminescent reagent according to claim 1 wherein the metal of the metal chelate is a rare earth metal or a metal of Group VIII.

14. The chemiluminescent reagent according to claim 1 wherein the metal is selected from the group consisting of europium, terbium, dysprosium, samarium osmium and ruthenium.

15. The chemiluminescent reagent according to claim 1 wherein the metal chelate comprises a chelating agent selected from the group consisting of NHA, BHHT, BHHCT, DPP, TTA, NPPTA, NTA, TOPO, TPPO, BFTA, 2,2-dimethyl-4-perfluorobutyoyl-3-butanone (fod), 2,2′

-dipyridyl (bpy), bipyridylcarboxylic acid, aza crown ethers, aza cryptands and trioctylphosphine oxide and derivatives thereof.

16. The chemiluminescent reagent according to claim 1 wherein the olefinic compound is selected from the group consisting of thioxenes, dioxenes, enol ethers, enamines, 9-alkylidenexanthans, 9-alkylidene-N-alkylacridans, aryl vinyl ethers, arylimidazoles and lucigenin and derivatives thereof.

17. The chemiluminescent reagent according to claim 1 wherein the metal is europium, the chelating agent is BHHCT and the olefinic compound is a carboxyl derivative of thioxene.

18. A method for determining the presence and/or amount of an analyte in a sample suspected of containing the analyte, the method comprising:

(a) providing in combination in a medium;

(i) the sample,

(ii) a chemiluminescent reagent, the reagent being non-particulate and comprising a binding partner for the analyte linked to a chemiluminescent composition comprising a metal chelate and an olefinic compound, and

(iii) a sensitizer reagent capable of generating singlet oxygen,

(b) subjecting the combination to conditions for binding of the analyte to the specific binding pair member for the analyte, and

(c) activating the sensitizer and detecting the amount of luminescence generated by the chemiluminescent composition, the amount of luminescence being related to the amount of the analyte in the sample.

19. The method according to claim 18 wherein the binding partner for the analyte is covalently bound to the chemiluminescent composition by a linking group and wherein the linking group comprises a hydrophilic macromolecule.

20. The method according to claim 18 wherein the binding partner for the analyte is non-covalently bound to the chemiluminescent composition by a specific binding pair wherein one member of the specific binding pair is linked to the binding partner for the analyte and the other member of the specific binding pair is linked to the chemiluminescent composition by a linking group wherein the linking group comprises a hydrophilic macromolecule.

21. The method of claim 18 wherein a binding partner for the analyte is associated with the sensitizer reagent and the sensitizer reagent comprises a particle.