Method of immobilizing a protein or molecule via a mutant dehalogenase that is bound to an immobilized dehalogenase substrate and linked directly or indirectly to the protein or molecule

US7888086B2 · kind B2 · utility

Assignee

• Promega Corporation · US

Inventors

• Aldis Darzins · Verona, US
• Lance P. Encell · Fitchburg, US
• Tonny Johnson · Madison, US
• Dieter Klaubert · Arroyo Grande, US
• Georgyi V. Los · Madison, US
• Mark McDougall · Bethlehem, US
• Keith V. Wood · Verona, US
• Monika G. Wood · Verona, US
• Chad Zimprich · Stoughton, US

Key dates

• Filing date: Jul 24, 2008
• Grant date: Feb 15, 2011
• Expiry date: Jan 31, 2029

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y308/01005
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.