Mutant protease biosensors with enhanced detection characteristics

US8735559B2 · kind B2 · utility

Assignee

• Promega Corporation · US

Inventors

• Brock Binkowski · Sauk City, US
• Braeden Butler · Madison, US
• Lance P. Encell · Fitchburg, US
• Frank Fan · Verona, US
• Brad Hook · Baraboo, US
• Paul Otto · Madison, US
• Gediminas Vidugiris · Fitchburg, US
• Susan Wigdal · Madison, US
• Kristopher Zimmerman · Madison, US

Key dates

• Filing date: May 11, 2011
• Grant date: May 27, 2014
• Expiry date: Jul 11, 2031

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y113/12007
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.