Compositions for the detection of biological molecules using a two particle complex

• US 8,871,917 B2
• Filed: 04/26/2012
• Issued: 10/28/2014
• Est. Priority Date: 06/23/2004
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Bard, Allen J., Miao, Wujian
• Primary Examiner(s): Chunduru, Prabha
• Application Number:
  US13/457,086
• Publication Number:
  US 20120329173A1
• Time in Patent Office: 915 Days
• Field of Search: None
• US Class Current: 536/22.1
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2563/103 luminescence
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: C12Q 2565/515 characterised by the interaction between or sequential use of two or more arrays
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Methods, compositions and kits for detecting analytes of interest in a sample using electrogenerated chemiluminescence are provided. Compositions comprising at least one solid support that entraps or contains an electrogenerated chemiluminescent moiety also provided.

Claims

1. A composition for detecting an analyte of interest in a sample comprising:

(A)_k(B)_u,(C),(D)_x

wherein A is an electrogenerated chemiluminescent (ECL) moiety which can be induced to repeatedly emit electromagnetic radiation by direct exposure to an electrochemical energy source; and

the ECL moiety is soluble in organic solvent and insoluble in aqueous solvent;

B is a first carrier comprising a plurality of the ECL moieties entrapped within its interior; and

B is either linked to the analyte of interest or linked to a first specific binding partner of the analyte of interest; and

the first carrier is a synthetic organic polymer bead;

C is the sample which may contain the analyte of interest; and

D is a second carrier is either linked to the analyte of interest or linked to a second specific binding partner of the analyte of interest;

wherein the second carrier is a magnetizable bead;

wherein k, u, and x are each an integer equal to or greater than 1; and

the first and second carriers are not both linked to the analyte of interest.

3. The composition of claim 1, wherein B is linked to the first specific binding partner;

and D is linked to the second specific binding partner.

6. The composition of claim 1, wherein the first carrier is a polystyrene bead.

11. The composition of claim 1, wherein said composition further comprises an amine ECL coreactant;

the analyte of interest is a protein;

the first specific binding partner is an antibody, a part of an antibody, or a binding protein; and

the second specific binding partner is an antibody, a part of an antibody, or a binding protein; and

the ECL moiety comprises three bidentate ligands and a metal ion selected from osmium or ruthenium.

12. The composition of claim 1, further comprising an amine ECL coreactant;

wherein the first carrier is a polystyrene bead;

the amine ECL coreactant comprises tripropylamine; and

the ECL moiety comprises bis(2,2′

-bipyridyl)-ruthenium(II) or tris(2,2′

-bipyridyl)ruthenium(II).

13. The composition of claim 1, wherein the ECL moieties comprise ruthenium and three bidentate ligands.

17. The composition of claim 1 wherein the analyte of interest is a protein;

the first specific binding partner is an antibody, a part of an antibody, or a binding protein; and

each ECL moiety comprises ruthenium and three bidentate ligands.

18. The composition of claim 1 wherein the synthetic organic polymethacrylate bead comprises a compound chosen from polystyrene, polyacrylic, nylon, polymethacrylate, polyacrylamide, polyacrylonitrile, PVDF, poly(lactide-co-ethylene glycol), and polyolefins.

19. The composition of claim 1 wherein the analyte of interest is a nucleic acid;

the first specific binding partner is a nucleic acid; and

each ECL moiety comprises ruthenium and three bidentate ligands.

20. The composition of claim 1 wherein the analyte of interest is an antigen;

the synthetic organic polymer bead is a polystyrene bead;

the first specific binding partner is an antibody or a part of an antibody; and

each ECL moiety comprises bis(2,2′

-bipyridyl)ruthenium(II) or tris(2,2′

-bipyridyl)ruthenium(II).

21. The composition of claim 20 wherein each ECL moiety comprises Ru(bpy)₃[B(C₆F₅)₄]₂.

2. A kit for detecting an analyte of interest in a sample comprising:

at least one first carrier comprising a plurality of electrogenerated chemiluminescent (ECL) moieties entrapped within its interior;

wherein the first carrier is either linked to the analyte of interest or linked to a first specific binding partner of the analyte of interest; and

the first carrier is a synthetic organic polymer bead;

at least one second carrier, which is either linked to the analyte of interest or linked to a second specific binding partner of the analyte of interest;

wherein the second carrier is a magnetizable bead;

provided that the first carrier and the second carrier are not both linked to the analyte of interest;

wherein the ECL moieties can be induced to repeatedly emit electromagnetic radiation by direct exposure to an electrochemical energy source; and

the ECL moieties are soluble in organic solvent and insoluble in aqueous solvent.

4. The kit of claim 2, wherein the analyte of interest is a protein;

the first specific binding partner is an antibody, a part of an antibody, or a binding protein; and

the second specific binding partner is an antibody, is part of an antibody, or a binding protein.

5. The kit of claim 2, wherein the analyte of Interest is a nucleic acid;

the first specific binding partner is to nucleic acid; and

the second specific binding partner is a nucleic acid.

7. The kit of claim 2, wherein the first carrier is a polystyrene bead.

8. The kit of claim 2, wherein the synthetic organic polymer bead comprises a compound chosen from polystyrene, polyacrylic, nylon, polymethacrylate, polyacrylamide, polyacrylonitrile, PVDF, poly(lactide-co-ethylene glycol), and polyolefins.

9. The kit of claim 2, wherein the ECL moiety comprises a metal ion selected from osmium or ruthenium.

10. The kit of claim 2, wherein the ECL moiety comprises bis(2,2′

-bipyridyl)-ruthenium(II) or tris(2,2′

-bipyridyl)ruthenium(II).

14. The kit of claim 2, wherein the ECL moieties comprise bis(2,2′

-bipyridyl)ruthenium(II) a tris(2,2′

-bipyridyl)ruthenium(II).

15. The kit of claim 14, wherein the ECL moieties comprise Ru(bpy)₃[B(C₆F₅)₄]₂.

16. The kit of claim 2, wherein the analyte of interest is a protein;

the first specific binding partner is an antibody, a part of an antibody, or a binding protein; and

the second specific binding partner is an antibody, a part of an antibody, or a binding protein; and

each ECL moiety comprises ruthenium and three bidentate ligands.

22. The kit of claim 2, wherein the analyte of interest is an antigen;

the first carrier is a polystyrene bead;

the first specific binding partner is an antibody or a part of an antibody;

the second specific binding partner is an antibody or a part of an antibody; and

each ECL moiety comprises bis(2,2′

-bipyridyl)ruthenium(II) or tris(2,2′

-bipyridyl)ruthenium(II).

23. The kit of claim 22, further comprising an amine ECL coreactant.

24. The kit of claim 23, wherein each ECL moiety comprises Ru(bpy)₃[B(C₆F₅)₄]₂;

and the amine ECL coreactant comprises tripropylamine.