Metallic solid supports modified with nucleic acids

• US 6,265,155 B1
• Filed: 02/28/1997
• Issued: 07/24/2001
• Est. Priority Date: 06/07/1995
• Status: Expired due to Term

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Meade, Thomas J., Kayyem, Jon F.
• Primary Examiner(s): Houtteman, Scott W.
• Application Number:
  US08/808,750
• Time in Patent Office: 1,607 Days
• Field of Search: 435/6, 435/5, 435/91.1, 536/23.1, 536/24.3, 536/24.32, 536/24.33, 422/69
• US Class Current: 435/6.11
• 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. An array of nucleic acids comprising a solid support having at plurality of regions, each region comprising a metallic surface comprising a self-assembled mixed monolayer comprising:

a) blocking moieties, having at least a first end attached to said metallic surface and at least a second end comprising a terminal group, wherein said blocking moieties shield nucleic acids from said metallic surface; and

b) at least one modified nucleic acid, comprising a nucleic acid and a linker moeity having a first and second end, wherein said first end of said linker is attached to said metallic surface and said second end is attached to said nucleic acid;

wherein at least two different regions comprise different nucleic acids.

2. An array according to claim 1 wherein said first end of said blocking moieties is attached to said metallic surface via a sulfur linkage.

3. An array according to claim 1 wherein each metallic surface comprises gold.

4. An array according to claim 1 wherein said blocking moieties have the formula:

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.

5. An array according to claim 4 wherein n is ≧

6.

6. An array according to claim 1 wherein said modified nucleic acids have the formula:

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.

7. An array according to claim 6 wherein n is ≧

6.

8. An array according to claim 1 wherein said blocking moiety comprises a phosphorus-containing moiety.

9. An array according to claim 1 wherein said nucleic acid is attached to said linker at a 2′

position of a ribose.

10. An array according to claim 1 wherein said nucleic acid is attached to said linker at a 3′

position of a ribose.

11. An array according to claim 1 wherein said nucleic acid is attached to said linker at a base of said nucleic acid.

12. An array according to claim 1 wherein said nucleic acid is attached to said linker at a phosphate linkage of said nucleic acid.

13. An array according to claim 1 wherein said solid support is glass.

14. An array according to claim 1 wherein said solid support is plastic.

15. A method of hybridizing a probe nucleic acid to a target nuclici acid, said method comprising adding a target nucleic acid to an array of probe nucleic acids comprising a solid support having at plurality of regions, each region comprising a metallic surface comprising a self-assembled mixed monolayer comprising:

a) blocking moieties, having at least a first end attached to said metallic surface and at least a second end comprising a tenninal group, wherein said blocking moieties shield nucleic acids from said metallic surface; and

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

wherein at least two different regions comprise different probe nucleic acids;

under conditions wherein at least one of said probe nucleic acids and said target nucleic acid will hybridize to form a hybridization complex.

16. A method according to claim 15 wherein said target nucleic acid is labelled.

17. A method according to claim 16 wherein said target nucleic acid is labelled with a fluorescent dye.