Nanoparticles having oligonucleotides attached thereto and uses therefor

• US 6,361,944 B1
• Filed: 06/25/1999
• Issued: 03/26/2002
• Est. Priority Date: 07/29/1996
• Status: Expired due to Term

• Current Assignee: Nanosphere Incorporated (DiaSorin S.p.A.), Northwestern University
• Original Assignee: Nanosphere Incorporated (DiaSorin S.p.A.)
• Inventors: Mucic, Robert C., Mirkin, Chad A., Storhoff, James J., Elghanian, Robert, Letsinger, Robert L.
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US09/344,667
• Time in Patent Office: 1,005 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 536/22.1, 536/23.1, 536/24.3, 536/24.33
• US Class Current: 435/6.11
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00603 Making arrays on substantially continuous surfaces
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B82B 1/00 Nanostructures formed by manipulation of individual atoms or molecules, or limited collections of atoms or molecules as discrete units
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• 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 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• 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 2525/161 incorporating target specific and non-target specific sites
• CPC Class Codes: C12Q 2537/125 : Sandwich assay format
• CPC Class Codes: C12Q 2537/143 : Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2563/143 : Magnetism, e.g. magnetic label
• CPC Class Codes: C12Q 2563/155 : Particles of a defined size, e.g. nanoparticles
• CPC Class Codes: C12Q 2565/625 : being a nucleic acid test strip device, e.g. dipsticks, strips, tapes, CD plates
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

The invention provides methods of detecting a nucleic acid. The methods comprise contacting the nucleic acid with one or more types of particles having oligonucleotides attached thereto. In one embodiment of the method, the oligonucleotides are attached to nanoparticles and have sequences complementary to portions of the sequence of the nucleic acid. A detectable change (preferably a color change) is brought about as a result of the hybridization of the oligonucleotides on the nanoparticles to the nucleic acid. The invention also provides compositions and kits comprising particles. The invention further provides nanomaterials and nanostructures comprising nanoparticles and methods of nanofabrication utilizing the nanoparticles. Finally, the invention provides a method of separating a selected nucleic acid from other nucleic acids.

Claims

1. A method for detecting a nucleic acid having a first and second portion comprising:

(a) providing a substrate having attached thereto oligonucleotides complementary to the first portion of the nucleic acid;

(b) providing gold nanoparticles having bound thereto by way of sulfur linkages oligonucleotides complementary to the second portion of the nucleic acid;

wherein the oligonucleotides complementary to the second portion of the nucleic acid to be detected are bound to the gold nanoparticle in an aging process comprising fire binding oligonucleotides to the gold nanoparticle in water and then binding additional oligonucleotides to the gold nanoparticle in a salt solution;

(c) contacting the substrate and gold nanoparticles provided in (a) and (b), respectively, with the nucleic acid under hybridizing conditions to bind the nucleic acid to the substrate and the gold nanoparticles; and

(d) detecting the gold nanoparticles bound to the nucleic acid bound to the substrate.

2. The method of claim 1 wherein the nucleic acid is contacted with the substrate provided in (a) prior to contacting it with the gold nanoparticles provided in (b).

3. The method of claim 1 wherein the nucleic acid is contacted with the gold nanoparticles provided in (b) prior to contacting it with the substrate provided in (a).

4. The method according to claim 1 wherein the gold nanoparticles are aged in a salt solution for about 40 hours to increase the density of oligonucleotide bound to the gold particles through sulfur linkages.

5. The method of claim 1 wherein the gold nanoparticle comprises oligonucleotides complementary to oligonucleotides on second gold nanoparticles and form an aggregate with the second gold nanoparticles.

6. The method of claim 1 wherein the gold nanoparticle is detected with a silver stain.

8. The method according to claim 1 wherein the nucleic acid is RNA or DNA.

9. The method according to claim 1 wherein the nucleic acid is human, bacterial or viral or fungal origin.

10. The method according to claim 1 wherein the nucleic acid is a gene associated with a disease.

11. The method of claim 1 wherein the nucleic acid is a synthetic DNA, a synthetic RNA, a structurally-modified natural or synthetic RNA, or a structurally-modified natural or synthetic DNA.

12. The method of claim 1 wherein the nucleic acid is a product of a polymerase chain reaction amplification.

7. The method of 5 wherein the oligonucleotide is detected with a silver stain.