Site-specific immobilization of gold binding polypeptide on gold nanoparticle-coated graphene sheet for biosensor application

Abstract

The effective and strong immobilization of enzymes on solid surfaces is required for current biological applications, such as microchips, biofuel cells, and biosensors. Gold-binding polypeptide (GBP), a genetically designed peptide, possesses unique and specific interactions with a gold surface, resulting in improved enzyme stability and activity. Herein we demonstrated an immobilization method for biosensor applications through site-specific interactions between GBP-fused organophosphorus hydrolase (GBP-OPH) and gold nanoparticle-coated chemically modified graphene (Au-CMG), showing enhanced sensing capability. A flow injection biosensor was fabricated by using GBP-OPH/Au-CMG to detect paraoxons, a model pesticide, showing higher sensitivity, lower detection limit and better operating stability compared that of OPH/Au-CMG. This strategy, which integrates biotic and abiotic moieties through site-specific interactions, has a great potential for use in biosensing and bioconversion process.