Title: Development of sensitive FRET sensors and methods of using the same

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.

Inventors:

Frommer, Wolf B.; Okumoto, Sakiko; Looger, Loren; Fehr, Marcus

Issue Date:

Tue Sep 10 00:00:00 EDT 2013

Research Org.:

Carnegie Inst. of Washington, Washington, DC (United States)

Sponsoring Org.:

USDOE

OSTI Identifier:

1531872

Patent Number(s):

8530633

Application Number:

11/665,339

Assignee:

Carnegie Institution of Washington (Washington, DC)

Patent Classifications (CPCs):

C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

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C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES

C07K14/245 - Escherichia

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/6818 - involving interaction of two or more labels, e.g. resonant energy transfer

G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES

G01N2800/2814 - Dementia
G01N2800/2835 - Movement disorders, e.g. Parkinson, Huntington, Tourette
G01N2800/2857 - Seizure disorders
G01N2800/2871 - Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
G01N33/542 - with steric inhibition or signal modification, e.g. fluorescent quenching
G01N33/566 - using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds}

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DOE Contract Number:  

FG02-04ER15542

Resource Type:

Patent

Resource Relation:

Patent File Date: 2005-10-14

Country of Publication:

United States

Language:

English