Nucleic Acid Ligand Diagnostic Biochip

摘要

A nucleic acid ligand “biochip” is disclosed, consisting of a solid support to which one or more specific nucleic acid ligands is attached in a spatially defined manner. Each nucleic acid ligand binds specifically and avidly to a particular target molecule contained within a test mixture, such as a bodily fluid. The target molecules include, but are not limited to, proteins (cellular, viral, bacterial, etc.) hormones, sugars, metabolic byproducts, cofactor, and intermediates, drugs, and toxins. Contacting the test mixture with the biochip leads to the binding of a target molecule to its cognate nucleic acid ligand. The biochip may then be contacted with a reagent(s) that reacts covalently with proteins and not with nucleic acids. Each protein target in the test mixture may then detected by detecting the presence of the reagent at the appropriate address on the biochip.

主权项

1. A method for detecting a target molecule that may be present in a test mixture, the method comprising:
(a) contacting a test mixture with a plurality of nucleic acid ligands, each said nucleic acid ligand comprising a first hybridization sequence that is unique to the nucleic acid ligand, and having a specific affinity for a target molecule, wherein a nucleic acid ligand-target complex is formed if said target molecule is present in said test mixture, and wherein the target is a protein, peptide, carbohydrate, lipid, virus, drug, cell or tissue, wherein said target molecule is a three dimensional target structure other than a polynucleotide that binds to said nucleic acid ligand through a mechanism which depends on Watson/Crick base pairing or triple helix binding; (b) providing a plurality of second hybridization sequences immobilized on a solid support, each said second hybridization sequence being complementary to corresponding said unique first hybridization sequence; (c) contacting said solid support comprising said plurality of complementary second hybridization sequences with said nucleic acid ligand-target complexes, such that each said unique first hybridization sequence is permitted to interact with said complementary second hybridization sequence; (d) at any point prior to (c), contacting said nucleic acid ligand with a detectable molecule; and (e) detecting said target molecule on said surface by detecting said detectable molecule.