US 2006/0234279 A1 · Screening using polarization anisotropy in FRET emissions

Abstract

Methods and apparatus are described for detecting specific binding between first and second chemical entities. The first chemical entity in association with a first fluorophore is immobilized. The second chemical entity is allowed to bind with the immobilized first chemical entity. The second chemical entity is or becomes coupled to a second fluorophore, which forms a FRET pair with the first fluorophore. The bound chemical entities are exposed to radiation at an excitation frequency for either the first or the second fluorophore, and polarization anisotropy of a FRET fluorescent signal from the bound chemical entities is measured to detect specific binding between the first and second chemical entities. Techniques are also disclosed for detecting whether a FRET interaction is occurring between a first chemical entity including a donor fluorophore and a second chemical entity including an acceptor fluorophore, using simultaneous anisotropy measurements at the wavelengths of the donor and acceptor fluorophores.

Claims37

Dependent claims

A method for detecting specific binding between a first chemical entity and a second chemical entity, the method comprising:

immobilizing the first chemical entity in association with a first fluorophore;
allowing the second chemical entity to bind with the immobilized first chemical entity, wherein the second chemical entity is or becomes coupled to a second fluorophore, which forms a FRET pair with the first fluorophore;
exposing the bound chemical entities to radiation at an excitation frequency for either the first or the second fluorophore;
and
measuring polarization anisotropy of a FRET fluorescent signal from the bound chemical entities to detect specific binding between the first and second chemical entities.

2 more elements

Dependencies are inferred from the claim text.

Screening using polarization anisotropy in FRET emissions

Life of this patent

Screening using polarization anisotropy in FRET emissions

Life of this patent

Abstract

Claims37