Graphitic nanotubes in luminescence assays

• US 5,866,434 A
• Filed: 03/06/1996
• Issued: 02/02/1999
• Est. Priority Date: 12/08/1994
• Status: Expired due to Term

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Fischer, Alan, Hoch, Robert, Lu, Ming, Martin, Mark T., Leland, Jonathan K., Massey, Richard J., Dong, Liwen, Jameison, Fabian, Liang, Pam
• Primary Examiner(s): CEPERLEY, MARY
• Application Number:
  US08/611,347
• Time in Patent Office: 1,063 Days
• Field of Search: 435/7.4, 435/7.5, 435/24, 435/26, 435/6, 435/7.92, 435/7.93, 435/7.94, 435/176, 435/182, 436/524, 436/526, 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, said assay-performance also being linked to a label compound capable of being induced to luminesce.

2. A nanotube according to claim 1, wherein said nanotube is graphitic and said luminescence is electrochemiluminescence.

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

(i) a graphitic nanotube containing a functional group and

(ii) an assay-performance-substance linked to said functional group, said assay-performance-substance being capable of binding to the analyte-of-interest.

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

(i) a graphitic nanotube containing a functional group and

(ii) an assay-performance-substance linked to said functional group, said assay-performance-substance being bound to the analyte of interest.

5. A composition according to claim 4, further comprising a second assay-performance-substance bound to the analyte, said second assay-performance-substance being linked to a label compound capable of being induced to luminesce.

7. A composition according to claim 5, wherein said assay-performance-substance contains at least one substance selected from the group consisting of

(i) added analyte of interest or added analogue of said analyte;

(ii) a binding partner of said analyte or a binding partner of said analogue; and

(iii) a reactive component capable of binding with (i) or (ii).

6. 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 and

(b) an assay-performance-substance linked to said functional group wherein said assay-performance-substance contains at least one substance selected from the group consisting of

(i) added analyte of interest or added analogue of said analyte;

(ii) a binding partner of said analyte or a binding partner of an analogue of said analyte; and

(iii) a reactive component capable of binding with (i) or (ii).

8. 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 first assay-performance-substance linked to a label compound capable of being induced to luminesce, and

(iii) a plurality of functionalized graphitic nanotubes bound to a second assay-performance-substance;

(b) incubating said composition to form a complex which includes said graphitic nanotube bound to said second assay-performance-substance and said label compound bound to said first assay-performance-substance;

(c) collecting said complex in a measurement zone;

(d) inducing the label compound in said complex to luminesce, and

(e) measuring the emitted luminescence to measure the presence of the analyte of interest in the sample.

9. A method as recited in claim 8 wherein said complex is collected on the surface of means for inducing luminescence and measuring said luminescence.

10. A method as recited in claim 9 based upon measurement of electrochemiluminescence wherein said complex is collected at an electrode surface.

11. A method for performing a binding assay for an analyte of interest present in a sample based upon measurement of electrochemiluminescence at an electrode surface comprising the steps:

(a) forming a composition containing

(i) said sample

(ii) a first assay-performance-substance linked to a label compound capable of being induced to electrochemiluminesce, and

(iii) a plurality of functionalized graphitic nanotubes bound to a second assay-performance-substance;

(b) incubating said composition to form a complex which includes said graphitic nanotube bound to said second assay-performance-substance and said label compound bound to said first assay-performance-substance;

(c) collecting said complex;

(d) causing said collected complex to come in contact with an electrode surface and inducing the label compound in said complex to luminesce by impressing a voltage on said electrode; and

(e) measuring the emitted luminescence at the electrode surface to measure the presence of the analyte of interest in the sample.

12. A method for performing a binding assay for an analyte of interest present in a sample based upon measurement of electrochemiluminescence at an electrode surface comprising the steps:

(a) forming a composition containing

(i) said sample

(ii) A first assay-performance-substance linked to a label compound capable of being induced to electrochemiluminesce, and

(iii) a plurality of magnetically responsive suspended graphitic nanotubes bound to a second assay-performance-substance;

(b) incubating said composition to form a complex which includes a graphitic nanotube and said label compound;

(c) collecting said complex by imposition of a magnetic field on said graphitic nanotubes;

(d) causing said collected complex to come in contact with an electrode surface and inducing the label compound in said complex to luminescence by imposing a voltage on said electrode; and

(e) measuring the emitted luminescence at the electrode surface to measure the presence of the analyte of interest in the sample.

13. A method as recited in claim 12 wherein the imposition of said magnetic field causes said complex to collect at the surface of said electrode.

14. A composition of matter for use as a reagent in a microparticulate-based binding assay comprising functionalized graphitic nanotubes bound to an assay-performance-substance and at least one other component selected from the group consisting of:

(a) electrolyte;

(b) label compound containing an ECL moiety;

(c) analyte of interest or an analog of the analyte of interest;

(d) a binding partner of the analyte of interest or of its analog;

(e) a reactive component capable of reacting with (c) or (d);

(f) a reductant; and

(g) an electrochemiluminescent-reaction enhancer;

provided, however, that no two components contained within any reagent composition are reactive with one another during storage so as to impair their function in the intended assay.

15. A reagent as recited in claim 14 containing magnetically responsive graphitic nanotubes.

16. An assay reagent for an assay based upon a binding reaction and the measurement of an electrochemiluminescent phenomenon comprising:

(a) an electrolyte;

(b) a plurality of magnetically responsive functionalized graphitic nanotubes bound to a first assay-performance-substance; and

(c) a second assay-performance-substance bound to a chemical moiety having electrochemiluminescent properties.

17. A method for performing an assay for an analyte of interest present in a sample, which analyte is an enzyme, comprising the steps of:

(a) forming a composition containing

(i) said sample, and

(ii) a functionalized graphitic nanotube bound to a component which is bound to a label compound capable of being induced to luminesce, wherein said component is a substrate of the analyte of interest;

(b) incubating said composition under conditions to permit said analyte to cleave said component, whereby a label sub-component free from said nanotube results from said cleavage;

(c) separating said graphitic nanotube from said composition;

(d) inducing the free label sub-component to luminesce; and

(e) measuring the emitted luminescence to measure the presence of the analyte of interest in the sample.

18. A graphitic nanotube linked to at least two components selected from the group consisting of (i) an electrochemiluminescent label, (ii) an enzyme and (iii) an enzyme cofactor.

19. A graphitic nanotube linked to an enzyme biosensor, wherein said enzyme biosensor includes a label compound capable of being induced to electrochemiluminesce.