High‐Throughput Electrochemical Screening Assay for Free and Immobilized Oxidases: Electrochemiluminescence and Intermittent Pulse Amperometry

Abstract

Analysis of a large number of samples by electrochemical methods for rapid optimization of amperometric biosensors or screening applications is still a challenge. To overcome this limitation, a system with 96 screen‐printed electrodes was developed to assay H2O2 produced by free and immobilized galactose oxidase as a model for oxidases. Detection was based on two electrochemical methods: intermittent pulse amperometry (IPA) and electrochemiluminescence (ECL). This 96‐well electrochemical device allowed rapid optimization of the parameters in half a day, regardless of the method used (i.e., IPA or ECL). We demonstrate that ECL is more sensitive for the detection of H2O2 (limit of detection of 1 μm for ECL vs. 10 μm for IPA) with lower interelectrode variability (1 vs. 19 %). Rapid determination of sensitivities, linear ranges, and limits of detection of galactose oxidase substrates (e.g., dihydroxyacetone and l‐erythrulose) by using free or immobilized enzyme was conducted in only 50 min by the mean of 5 different sets of 96 screen‐printed electrodes.

Highly Sensitive Electrochemiluminescence Assay for Cardiac Troponin I and Adenosine Triphosphate by using Supersandwich Amplification and Bifunctional Aptamer

“…[1] A variety of assay has been developed for detection of multiple analytes, such as proteins, nucleic acids, tumors, toxins, and small molecules. [2] Most detection of multiple analytes is usually accomplished by constructing sensor arrays with each component in the array sensitive to a specific analyte or to several analytes. The complexity of sensor array made the development of simple, sensitive and specific multicomponent sensing system for the detection of multiple targets are urgent needed.…”

High-Throughput Electrochemistry: State of the Art, Challenges, and Perspective

“…The arrays generally consist of 96-well plates containing two carbon electrodes that have been screen-printed onto a printed circuit board. In addition to biomolecule detection, screen-printed multielectrode arrays have been adapted for high-throughput screening of enzyme libraries generated from directed evolution. , In these systems, the electrodes in each well are covered with 35–200 μL of electrolyte, and the enzymatic activity is determined using intermittent pulse amperometry. This many electrodes, many electrolytes approach has been used to select enzymes for biofuel cells and synthetic biology as well as the screening of enzyme inhibitors .…”

“…In addition to biomolecule detection, screen-printed multielectrode arrays have been adapted for high-throughput screening of enzyme libraries generated from directed evolution. , In these systems, the electrodes in each well are covered with 35–200 μL of electrolyte, and the enzymatic activity is determined using intermittent pulse amperometry. This many electrodes, many electrolytes approach has been used to select enzymes for biofuel cells and synthetic biology as well as the screening of enzyme inhibitors . Screen-printed multielectrode arrays have also been developed in a three-electrode format to allow more precise control over the electrode potential .…”

Electrostatic Self‐Assembled Bracelet‐Like Au@Pt Nanoparticles: An Efficient Electrocatalyst for Highly Sensitive Non‐Enzymatic Hydrogen Peroxide Sensing

“…[3] Thus, it is also essential to seek an efficient method for highly sensitive and accurate detection of H 2 O 2 . Up to present, several methodshave been established for the detection of H 2 O 2 , including spectrophotometry, [4] fluorescence, [5] chemiluminescence, [6] electrochemical method. [7] Among these methods, the electrochemical sensing platform has been emphasized due to its low cost, simplicity, and excellent detection performance.…”