title: A 96-well electrochemical method for the screening of enzymatic activities.
abstract: The rapid electrochemical screening of enzyme activities in bioelectronics is still a challenging issue. In order to solve this problem, we propose to use a 96-well electrochemical assay. This system is composed of 96 screen-printed electrodes on a printed circuit board adapted from a commercial system (carbon is used as the working electrode and silver chloride as the counter/reference electrode). The associated device allows for the measurements on the 96 electrodes to be performed within a few seconds. In this work, we demonstrate the validity of the screening method with the commercial laccase from the fungus Trametes versicolor. The signal-to-noise ratio (S/N) is found to be the best way to analyze the electrochemical signals. The S/N follows a saturation-like mechanism with a dynamic linear range of two decades ranging from 0.5 to 75 ng of laccase (corresponding to enzymatic activities from 62 × 10(-6) to 9.37 × 10(-3) μmol min(-1)) and a sensitivity of 3027 μg(-1) at +100 mV versus Ag/AgCl. Laccase inhibitors (azide and fluoride anions), pH optima, and interfering molecules could also be identified within a few minutes.
authors: GEMBAS (Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires), ICBMS UMR 5246, Université Lyon 1, CNRS, INSA Lyon, Villeurbanne, France.
authors: Sofiène
authors: S
authors: Abdellaoui
authors: Alexandre
authors: A
authors: Noiriel
authors: Robert
authors: R
authors: Henkens
authors: Celia
authors: C
authors: Bonaventura
authors: Loïc J
authors: LJ
authors: Blum
authors: Bastien
authors: B
authors: Doumèche
mesh_terms: D055664:Electrochemical Techniques / Q000295:instrumentation*; D004566:Electrodes; D057075:Enzyme Assays / Q000295:instrumentation*; D004867:Equipment Design; D042845:Laccase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008958:Models, Molecular; D012680:Sensitivity and Specificity; D059629:Signal-To-Noise Ratio; D055454:Trametes / Q000201:enzymology*
chemical_list: D042845:Laccase