Nucleic acid ligand diagnostic biochip

• US 6,544,776 B1
• Filed: 08/14/2000
• Issued: 04/08/2003
• Est. Priority Date: 12/15/1997
• Status: Expired due to Term

• Current Assignee: SomaLogic Operating Co., Inc. (Somalogic Incorporated)
• Original Assignee: Somalogic Incorporated
• Inventors: Gold, Larry, Zichi, Dominic A., Jayasena, Sumedha, Drolet, Daniel, Gill, Stanley, Creighton, Steve
• Primary Examiner(s): Siew, Jeffrey
• Application Number:
  US09/581,465
• Time in Patent Office: 967 Days
• Field of Search: 435/6, 435/7.1, 435/91.1, 435/91.2, 536/22.1, 536/23.1, 536/24.3, 536/24.31, 536/24.32, 536/24.33
• US Class Current: 435/287.2
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• 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 1/6839 Triple helix formation or other higher order conformations in hybridisation assays
• CPC Class Codes: C12Q 2525/205 Aptamer
• CPC Class Codes: C12Q 2541/101 Selex
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: C12Q 2565/514 characterised by the use of the arrayed oligonucleotides as identifier tags, e.g. universal addressable array, anti-tag or tag complement array
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures
• CPC Class Codes: Y10S 977/924 using nanostructure as support of dna analysis

Abstract

A diagnostic biochip comprising a solid support to which one or more specific nucleic acid ligands is attached in a spatially defined manner is provided. Each nucleic acid ligand binds specifically and avidly to a particular target molecule contained within a test mixture, such as a bodily fluid. Also provided are methods for the preparation of nucleic acid ligand biochips. Further, methods for the use of the nucleic acid ligand biochip in diagnosis of a medical condition and quantitative detection of a target molecule are provided.

Claims

1. A biochip comprising an array of a plurality of biochip bound nucleic acid ligands, wherein a plurality of said nucleic acid ligands are specifically associated with a target molecule through non-Watson-Crick interactions.

2. The biochip of claim 1, wherein each of said nucleic acid ligands comprises a spectroscopically detectable label.

3. The biochip of claim 2, wherein the spectroscopically detectable label is selected from the group consisting of a fluorescent label, a luminescent label, an energy transfer label, and a phosphorescent label.

4. The biochip of claim 1, wherein each of said nucleic acid ligands comprises a photoreactive group.

5. The biochip of claim 1, wherein said target molecule is a protein.

6. The biochip of claim 1, wherein the association between the nucleic acid ligand and the target molecule is covalent.

7. The biochip of claim 1, further comprising a crosslinking agent, said crosslinking agent being covalently attached to the biochip, said each of said nucleic acid ligands being attached to the crosslinking agent.

8. The biochip of claim 1, wherein each of said biochip-bound nucleic acid ligands are bound to said biochip via a covalent bond, said covalent bond being formed by the reaction of a first reactive group on said each of said nucleic acid ligands, and a second reactive group on a surface of the biochip.

9. The biochip of claim 8, wherein said first reactive group is a thiol group, and said second reactive group is a thiol group.

10. The biochip of claim 8, wherein said first reactive group is a 3′

-hydroxyl group, and said second reactive group is a nitroveratryl group.

11. The biochip of claim 1, wherein each of said biochip-bound nucleic acid ligands are bound to said biochip via a non-covalent association, said non-covalent association being formed by the interaction of a first group on said each of said nucleic acid ligands, and a second group on a surface of the biochip.

12. The biochip of claim 11, wherein said first group is biotin, and said second group is streptavidin.

13. The biochip of claim 1, wherein the target molecule comprises a detectable label.

14. The biochip of claim 1, wherein the target molecule is associated with an antibody with a detectable label.

15. The biochip of claim 1, wherein the target molecule is detectable by an antibody sandwich assay.

16. The biochip of claim 1, wherein the target molecule is associated with a primary antibody, said primary antibody being associated with a molecule selected from the group consisting of a secondary antibody with a detectable label, a secondary antibody conjugated to a reporter enzyme, which enzyme catalyzes the production of a detectable compound, and protein A with a detectable label.

17. The biochip of claim 1, wherein the target molecule is associated with a second nucleic acid ligand selected from the group consisting of a nucleic acid ligand with a detectable label, a nucleic acid ligand with a fluorescent label, and a nucleic ligand conjugated to biotin.

18. The biochip of claim 1, wherein the target molecule is detectable by a competition assay.