title: Formation and characterization of glutamate dehydrogenase monolayers on silicon supports.
abstract: In this paper we have tested two different procedures (the "three-step" and the "four-step" procedures) for the covalent immobilization of glutamate dehydrogenase (GDH) onto silicon supports. Atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FT-IR), fluorescence spectroscopy and an enzymatic assay were used to probe the structure and activity of the immobilized enzyme. Our results demonstrate that coupling through the "three-step" procedure does not significantly affect either the fold pattern or the activity of the enzyme, suggesting that this method could be ideally suited to the development of high quality monolayers for use in enzyme-based planar biosensors.
authors.affiliation: National Nanotechnology Laboratory of INFM, c/o Department of Innovation Engineering, University of Lecce, Italy. laura.blasi@unile.it
authors.forename: L
authors.initials: L
authors.lastname: Blasi
authors.forename: L
authors.initials: L
authors.lastname: Longo
authors.forename: P P
authors.initials: PP
authors.lastname: Pompa
authors.forename: L
authors.initials: L
authors.lastname: Manna
authors.forename: G
authors.initials: G
authors.lastname: Ciccarella
authors.forename: G
authors.initials: G
authors.lastname: Vasapollo
authors.forename: R
authors.initials: R
authors.lastname: Cingolani
authors.forename: R
authors.initials: R
authors.lastname: Rinaldi
authors.forename: A
authors.initials: A
authors.lastname: Rizzello
authors.forename: R
authors.initials: R
authors.lastname: Acierno
authors.forename: C
authors.initials: C
authors.lastname: Storelli
authors.forename: M
authors.initials: M
authors.lastname: Maffia
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D004800:Enzymes, Immobilized; D005969:Glutamate Dehydrogenase; D018625:Microscopy, Atomic Force; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D012825:Silicon; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D004800:Enzymes, Immobilized; D009243:NAD; D005969:Glutamate Dehydrogenase; D012825:Silicon