Development of sensitive FRET sensors and methods of using the same

US8530633B2 · kind B2 · utility

Assignee

• Carnegie Institution of Washington · US

Inventors

• Wolf B. Frommer · Washington, US
• Sakiko Okumoto · Washington, US
• Loren Looger · Sterling, US
• Marcus Fehr · Washington, US

Key dates

• Filing date: Oct 14, 2005
• Grant date: Sep 10, 2013
• Expiry date: May 11, 2028

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N2800/2871
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Intramolecular biosensors are disclosed, including PBP-based biosensors, comprising a ligand binding domain fused to donor and fluorescent moieties that permit detection and measurement of Fluorescence Resonance Energy Transfer upon binding ligand. At least one of the donor and fluorescent moieties may be internally fused to the biosensor such that both ends of the internally fused fluorophore are fixed. In addition, methods of improving the sensitivity of terminally fused biosensors are provided. The biosensors of the invention are useful for the detection and quantification of ligands in vivo and in culture.