Methods and apparatus for improved luminescence assays using microparticles

• US 6,846,629 B2
• Filed: 06/29/2001
• Issued: 01/25/2005
• Est. Priority Date: 05/09/1995
• Status: Expired due to Term

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Wohlstadter, Jacob N., Wilbur, James L., Martin, Mark T., Sigal, George B., Gudibande, Satyanarayana
• Primary Examiner(s): Siew, Jeffrey
• Application Number:
  US09/896,962
• Publication Number:
  US 20030003460A1
• Time in Patent Office: 1,306 Days
• Field of Search: 435/6, 435/26, 435/5, 435/91.2, 435/515, 536/26.6, 536/24.3, 536/24.33, 536/24.32, 536/7.1, 536/7.9
• US Class Current: 435/6.11
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. 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 2537/125 Sandwich assay format
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: G01N 2458/30 Electrochemically active labels
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/552 Glass or silica
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/587 Nanoparticles

Abstract

Microparticles comprising a core and electrochemiluminescent moieties within the core and methods of using the same in assays.

Claims

1. A microparticle comprising a core and electrochemiluminescent moieties within said core, wherein said electrochemiluminescent moieties are metal-containing electrochemiluminescent moieties.

2. The microparticle of claim 1, wherein said electrochemiluminescent moieties are blended within said core.

3. The microparticle of claim 1, wherein said core is polymeric and said electrochemiluminescent moieties are blended within said core.

4. The microparticle of claim 1, wherein said core comprises plastic and said electrochemiluminescent moieties are blended with said plastic to form said core.

5. The microparticle of claim 1, wherein said electrochemiluminescent moieties are enclosed within said core.

6. The microparticle of claim 1, wherein said core is a liposome and said electrochemiluminescent moieties are enclosed within said liposome.

7. The microparticle of claim 1, wherein said core comprises pores and said electrochemiluminescent moieties are enclosed within said pores.

8. The microparticle of claim 1, wherein said electrochemiluminescent moieties comprises transition metals.

9. The microparticle of claim 1, wherein said electrochemiluminescent moieties comprises Ru, Os, or Re.

10. The microparticle of claim 1, wherein said core is electrically conductive.

11. The microparticle of claim 1, wherein said core comprises metal.

12. The microparticle of claim 1, wherein said core comprises gold, silver, platinum, palladium, zinc, iron, nickel, lead or copper.

13. The microparticle of claim 1, wherein said core comprises gold, silver, platinum, or palladium.

14. The microparticle of claim 1, wherein said core comprises gold.

15. The microparticle of claim 1, wherein said core comprises carbon.

16. The microparticle of claim 1, wherein said core comprises carbon black, graphitic nanotubes or fullerenes.

17. The microparticle of claim 1, further comprising an assay ligand.

18. The microparticle of claim 17, wherein said assay ligand is selected from the group consisting of proteins, nucleic acids, lipids, steroids, carbohydrates, porphyrins, alkaloids, nucleotides, nucleosides, amino acids, fatty acids, viruses, microorganisms, biological cells, and subcellular particles.

19. The microparticle of claim 17, wherein said assay ligand is selected from the group consisting of proteins and nucleic acids.

20. An assay composition comprising the microparticle of claim 1 and at least one assay component selected from the group consisting of electrochemiluminescence co-reactant and binding reagent.

22. The microparticle of claim 1, further comprising one or more copies of an assay-ligand immobilized on its surface.

25. The microparticle of claim 1, wherein said microparticle has a diameter greater than 5 nanometers.

26. The microparticle of claim 1, wherein said microparticle has a diameter between 5 nanometers and 10 micrometers.

27. The microparticle of claim 1, wherein said microparticle has a diameter between 20 nanometers and 200 micrometers.

28. The microparticle of claim 1, wherein said microparticles are spherical, oblong, rod-like or irregular in shape.

21. A method for conducting electrochemiluminescence measurements for a binding analyte-of-interest comprising the steps of:

(a) forming a complex comprising;

(i) said analyte,

(ii) a microparticle comprising a core and electrochemiluminescent moieties within said core and having one or more copies of an assay-ligand immobilized on its surface, wherein said electrochemiluminescent moieties are metal-containing electrochemiluminescent moieties; and

(iii) an assay-ligand immobilized on an electrode; and

(b) conducting an electrochemiluminescence measurement at said electrode in the presence of electrochemiluminescence reactants.

24. The method of claim 21, wherein said assay-ligand immobilized on said surface and/or said assay-ligand immobilized on said electrode are capable of specifically binding with said analyte of interest.

23. A microparticle comprising metal-containing electrochemiluminescent moieties within said microparticle.