Graphitic nanotubes in luminescence assays

• US 6,362,011 B1
• Filed: 02/02/1999
• Issued: 03/26/2002
• Est. Priority Date: 12/08/1994
• Status: Expired due to Term

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Lu, Ming, Leland, Jonathan K., Jameison, Fabian, Liang, Pam, Fischer, Alan, Hoch, Robert, Martin, Mark T., Massey, Richard J., Dong, Liwen
• Primary Examiner(s): CEPERLEY, MARY
• Application Number:
  US09/243,215
• Time in Patent Office: 1,148 Days
• Field of Search: 435/6, 435/7.4, 435/7.5, 435/7.92, 435/7.93, 435/7.94, 435/24, 435/26, 435/176, 435/182, 436/524, 436/535, 436/806, 530/391.1, 530/391.3, 530/391.5
• US Class Current: 436/526
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 40/00 Manufacture or treatment of nanostructures
• CPC Class Codes: C01B 2202/34 Length
• CPC Class Codes: C01B 2202/36 Diameter
• CPC Class Codes: C01B 32/174 Derivatisation; Solubilisation; Dispersion in solvents
• CPC Class Codes: C07K 17/06 attached to the carrier via a bridging agent
• CPC Class Codes: D01F 11/14 with organic compounds, e.g. macromolecular compounds
• CPC Class Codes: D01F 9/12 Carbon filaments; Apparatus specially adapted for the manufacture thereof
• CPC Class Codes: G01N 2030/8863 Fullerenes
• CPC Class Codes: G01N 21/69 specially adapted for fluids , e.g. molten metal
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G11C 13/025 using fullerenes, e.g. C60, or nanotubes, e.g. carbon or silicon nanotubes
• CPC Class Codes: H01M 4/60 of organic compounds
• CPC Class Codes: Y02E 60/10 Energy storage using batteries
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 977/737 : having a modified surface
• CPC Class Codes: Y10S 977/742 : Carbon nanotubes, CNTs

Abstract

Graphitic nanotubes, which include tubular fullerenes (commonly called “buckytubes”) and fibrils, which are functionalized by chemical substitution, are used as solid supports in electrogenerated chemiluminescence assays. The graphitic nanotubes are chemically modified with functional group biomolecules prior to use in an assay. Association of electrochemiluminescent ruthenium complexes with the functional group biomolecule-modified nanotubes permits detection of molecules including nucleic acids, antigens, enzymes, and enzyme substrates by multiple formats.

Claims

1. A nanotube comprising carbon to which is attached an assay performance substance linked to an electrochemiluminescence label compound, wherein said label compound comprises a rare earth metal or a transition metal.

31. The nanotube of claim 1, wherein said label compound comprises a metal atom selected from the group consisting of ruthenium, osmium and rhenium.

32. The nanotube of claim 1, wherein said nanotube is magnetically responsive.

33. The nanotube of claim 1, wherein said label compound comprises:

(a) a metal atom selected from the group consisting of ruthenium, osmium and rhenium; and

(b) a nitrogen-containing, heterocyclic, aromatic polydentate metal ligand.

34. The nanotube of claim 1, wherein said assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acid.

2. A composition for the detection of an analyte of interest present in a sample, which composition comprises:

(a) a graphitic nanotube containing a functional group,

(b) a first assay-performance substance linked to said functional group; and

(c) a second assay-performance substance linked to an electroluminescence label compound, said label compound comprising a rare earth metal atom or a transition metal atom;

wherein said first and second assay-performance substances are selected from the group consisting of;

(i) specific binding partners of said analyte of interest; and

(ii) added analyte of interest or added analogues of said analyte of interest that compete with said analyte of interest for binding to a binding partner of said analyte of interest.

6. The composition of claim 2, wherein said label compound comprises a metal atom selected from the group consisting of ruthenium, osmium and rhenium.

10. The composition of claim 2, wherein said label compound comprises:

(a) a metal atom selected from the group consisting of ruthenium, osmium and rhenium; and

(b) a nitrogen-containing, heterocyclic, aromatic polydentate metal ligand.

14. The composition of claim 2, wherein said first assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acid.

17. The composition of claim 2, wherein said second assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acid.

20. The composition of claim 2, wherein said first assay performance substance is linked to said nanotube via a linkage comprising a biotin-streptavidin interaction.

27. The composition of claim 2, wherein said nanotube is magnetically responsive.

35. The composition of claim 2, wherein said first assay-performance substance is covalently linked to said fictional group.

3. A method for performing an assay for an enzyme present in a sample, comprising the steps of:

(a) forming a composition containing;

(i) said sample, and

(ii) a substrate for said enzyme, wherein said substrate is linked to a functionalized graphitic nanotube and said substrate is also linked to an electrochemiluminescence label compound, said label compound comprising a rare earth metal atom or a transition metal atom;

(b) incubating said composition under conditions that permit said enzyme, if present, to act on said substrate;

(c) exposing said composition, or a fraction thereof, to conditions suitable for inducing said label to electrochemiluminescence; and

(d) measuring said electrochemiluminescence so as to assay for said enzyme present in said sample.

7. The method of claim 3, wherein said label compound comprises a metal atom selected from the group consisting of ruthenium, osmium and rhenium.

11. The method of claim 3, wherein said label compound comprises:

(a) a metal atom selected from the group consisting of ruthenium osmium and rhenium; and

(b) a nitrogen-containing, heterocyclic, aromatic polydentate metal ligand.

21. The method of claim 3, wherein said first assay performance substance is linked to said nanotube via a linkage comprising a biotin-streptavidin interaction.

23. The method of claim 3, further comprising the step of collecting said nanotube at an electrode surface.

25. The method of claim 3, wherein said enzyme cleaves said substrate so as to release said label compound from said nanotube.

26. The method of claim 25, further comprising the step of separating said free label from said nanotube and wherein the fraction containing said free label is exposed to the conditions suitable for inducing said label to electrochemiluminescence.

28. The method of claim 3, wherein said nanotube is magnetically responsive.

4. A method for performing a binding assay for an analyte of interest present in a sample, comprising the steps of:

(a) forming a composition containing;

(i) said sample;

(ii) a graphitic nanotube containing a functional group;

(iii) a first assay-performance substance linked to said functional group; and

(iv) a second assay-performance substance linked to an electrochemiluminescence label compound, said label compound comprising a rare earth metal atom or a transition metal atom;

wherein said first and second assay-performance substances are selected from the group consisting of;

(1) specific binding partners of said analyte of interest; and

(2) added analyte of interest or added analogues of said analyte of interest that compete with said analyte of interest for binding to a binding partner of said analyte of interest;

(b) fog a complex comprising said graphitic nanotube and said label compound;

(c) inducing said label compound to emit electrochemiluminescence; and

(d) detecting the emitted electrochemiluminescence to measure the presence of the analyte of interest in the sample.

8. The method of claim 4, wherein said label compound comprises a metal atom selected from the group consisting of ruthenium, osmium and rhenium.

12. The method of claim 4, wherein said label compound comprises:

(a) a metal atom selected from the group consisting of ruthenium, osmium and rhenium; and

(b) a nitrogen-containing, heterocyclic, aromatic polydentate metal ligand.

15. A The method of claim 4, wherein said first assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acid.

18. The method of claim 4, wherein said second assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acid.

24. The method of claim 4, further comprising the step of collecting said nanotube at an electrode surface.

29. The method of claim 4, wherein said nanotube is magnetically responsive.

5. A kit for conducting an electrochemiluminescence binding assay comprising in one or more containers:

(a) a graphitic nanotube linked to a first assay-performance substance; and

(b) a second assay-performance substance linked to an electroluminescence label compound, said label compound comprising a rare earth metal atom or a transition metal atom.

9. The kit of claim 5, wherein said label compound comprises a metal atom selected from the group consisting of ruthenium, osmium and rhenium.

13. The kit of claim 5, wherein said label compound comprises:

(a) a metal atom selected from the group consisting of ruthenium, osmium and rhenium; and

(b) a nitrogen-containing, heterocyclic, aromatic polydentate metal ligand.

16. The kit of claim 5, wherein said first assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acid.

19. The kit of claim 5, wherein said second assay-performance substance comprises an antibody, a protein receptor, avidin, streptavidin, protein A, protein G, biotin or a nucleic acids.

22. The kit of claim 5, wherein said first assay performance substance is linked to said nanotube via a linkage comprising a biotin-streptavidin interaction.

30. The kit of claim 5, wherein said nanotube is magnetically responsive.