title: Site-specific immobilization of a (His)6-tagged acetylcholinesterase on nickel nanoparticles for highly sensitive toxicity biosensors.
abstract: This paper reports site-specific affinity immobilization of (His)6-tagged acetylcholinesterase (AChE) onto Ni/NiO nanoparticles for the development of an electrochemical screen-printed biosensor for the detection of organophosphate pesticides. The method is based on the specific affinity binding of the His-tagged enzyme to oxidized nickel nanoparticle surfaces in the absence of metal chelators. This approach allows stable and oriented attachment of the enzyme onto the oxidized nickel through the external His residue in one-step procedure, allowing for fast and sensitive detection of paraoxon in the concentration range from 10(-8) to 10(-13) M. A detection limit of 10(-12) M for paraoxon was obtained after 20 min incubation. This method can be used as a generic approach for the immobilization of other His-tagged enzymes for the development of biosensors.
authors.affiliation: Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, NY 13699-5810, USA.
authors.forename: Mallikarjunarao
authors.initials: M
authors.lastname: Ganesana
authors.forename: Georges
authors.initials: G
authors.lastname: Istarnboulie
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Marty
authors.forename: Thierry
authors.initials: T
authors.lastname: Noguer
authors.forename: Silvana
authors.initials: S
authors.lastname: Andreescu
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D003217:Conductometry / Q000295:instrumentation*; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006632:Histamine / Q000737:chemistry*; D010755:Organophosphates / Q000032:analysis* / Q000737:chemistry; D010575:Pesticides / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D018675:Toxicity Tests / Q000295:instrumentation*
chemical_list: D004800:Enzymes, Immobilized; D010755:Organophosphates; D010575:Pesticides; D006632:Histamine; D000110:Acetylcholinesterase