US 6,670,132 B2 · Determining non-nucleic acid molecule binding to target by competition with nucleic acid ligand

Abstract

A competitive binding assay is used to determine whether a non-nucleic acid molecule from a library of non-nucleic acid molecules binds to a target. The non-nucleic acid molecule competes with a nucleic acid ligand for binding to the target which may be immobilized. Detecting displacement of nucleic acid ligand from a complex of the nucleic acid ligand and the target determines binding of the non-nucleic acid molecule to the target. The nucleic acid ligand may be immobilized and contacted with a target to form a complex. Adding a non-nucleic acid molecule to the complex displaces target from the complex, and detecting displacement of the target determines binding of the non-nucleic acid molecule to the target. Labeled nucleic acid ligand or labeled target displaced from or remaining in the complex can be detected for detecting displacement. Nucleic acid ligands that bind to the target are identified by the SELEX method. The assay provides a high throughput screening method for determining whether a non-nucleic acid molecule binds to a target.

Claims12

Dependent claims

A method for determining whether at least one non-nucleic acid molecule from a library of candidate non-nucleic acid molecules binds to a target, wherein said non-nucleic acid molecule has an affinity for said target within a defined concentration range, comprising:

a) providing a first nucleic acid ligand of said target, said nucleic acid ligand having a specific binding affinity for said target;
b) modifying said first nucleic acid ligand of said target to yield a second nucleic acid ligand having a reduced binding affinity for said target relative to said first nucleic acid ligand;
c) immobilizing a target to a solid support;
d) contacting said second nucleic acid ligand with said target wherein binding of the second nucleic acid ligand to the target occurs to form a complex;
e) adding a non-nucleic acid molecule to the complex wherein the non-nucleic acid molecule competes with the second nucleic acid ligand for binding to the target and disrupts the complex and liberates the second nucleic acid ligand;
and
f) detecting the liberation of the second nucleic acid ligand thereby determining that said non-nucleic acid molecule binds to said target with an affinity within said defined concentration range.

4 more elements

a) providing a first nucleic acid ligand of said target, said first nucleic acid ligand having a specific binding affinity for said target;
b) modifying said first nucleic acid ligand of said target to yield a second nucleic acid ligand having a reduced binding affinity for said target relative to said first nucleic acid ligand;
c) immobilizing said second nucleic acid ligand to a solid support;
d) contacting said second nucleic acid ligand with a target wherein binding of the second nucleic acid ligand to the target occurs to form a complex;
e) adding a non-nucleic acid molecule to the complex wherein the non-nucleic acid molecule competes with the second nucleic acid ligand for binding to the target and disrupts the complex and liberates the target;
and
f) detecting the liberation of target thereby determining that said non-nucleic acid molecule binds to said target within said defined concentration range.

4 more elements

Dependencies are inferred from the claim text.

Determining non-nucleic acid molecule binding to target by competition with nucleic acid ligand

Life of this patent

Determining non-nucleic acid molecule binding to target by competition with nucleic acid ligand

Life of this patent

Abstract

Claims12