Metallic solid supports modified with nucleic acids

• US 20020034759A1
• Filed: 08/03/2001
• Published: 03/21/2002
• Est. Priority Date: 06/07/1995
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Meade, Thomas J., Kayyem, Jon F.
• Granted Patent: US 7,033,760 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• 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: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: C12Q 2565/1015 labels being on the same oligonucleotide
• CPC Class Codes: C12Q 2565/401 Signal amplification by chemical polymerisation
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode

Abstract

The present invention is directed to solid supports having metallic surfaces comprising blocking moieties and modified nucleic acids, which exhibit excellent characteristics in hybridization assays, in a stable, reproducible, rapid manner. In an additional aspect, the invention provides methods utilizing the solid supports to hybridize probe nucleic acid to target nucleic acid and methods for detecting the hybridization complex.

Claims

1. A solid support having a metallic surface comprising:

a) blocking moieties, having at least first and second ends, attached at said first end to said metallic surface;

b) at least one modified nucleic acid, comprising a linker moiety having a first and a second end, wherein said first end of said linker moiety is attached to said solid support and said second end is attached to a nucleic acid.

2. A solid support having a metallic surface comprising a covalently immobilized monolayer, wherein a subset of the molecules forming said monolayer are covalently linked to nucleic acid.

3. A solid support having a metallic surface comprising:

a) blocking moieties, comprising at least first and second ends, attached at said first end to said metallic surface via a sulfur linkage; and

b) modified nucleic acids, comprising a linker moiety having a first and a second end, wherein said first end of said linker moiety is attached to said metallic surface via a sulfur linkage, and said second end is attached to a nucleic acid.

4. A solid support having a metallic surface comprising:

a) blocking moieties having the formula comprising;

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said metallic surface;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups;

n is an integer from 3 to 50; and

X is a terminal group; and

b) modified nucleic acids having the formula comprising;

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said metallic surface;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups; and

n is an integer from 3 to 50.

6. A solid support according to claim 4 wherein said support comprises gold.

7. A solid support according to claim 4 wherein n is 16.

8. A solid support according to claim 4 wherein said blocking moieties are all the same.

9. A solid support according to claim 4 wherein at least two of said blocking moieties are different.

10. A solid support according to claim 4 wherein said linker moieties and said blocking moieties are the same.

11. A solid support according to claim 4 wherein at least one of said linker moieties and at least one of said blocking moieites are different.

13. The method according to claim 11, further comprising:

c) detecting said hybridization complex.

14. The method of claim 12, wherein said target nucleic acid is labelled and said detecting is done by detecting the presence of said label.

5. A solid support having a metallic surface comprising a mixed monolayer of:

a) blocking moieties having the formula comprising;

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said metallic surface;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups;

n is an integer from 3 to 50; and

X is a terminal group; and

b) modified nucleic acids having the formula comprising;

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said metallic surface;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups; and

n is an integer from 3 to 50.

12. A method of hybridizing probe nucleic acid to target nucleic acid, comprising adding target nucleic acid to a solid support having a metallic surface comprising:

a) blocking moieties having the formula comprising;

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said metallic surface;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups;

n is an integer from 3 to 50; and

X is a terminal group; and

b) modified nucleic acids having the formula comprising;

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said metallic surface;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups;

nucleic acid is said probe nucleic acid; and

n is an integer from 3 to 50.

under conditions where said probe nucleic acid and said target nucleic acid will hybridize to form a hybridization complex.