Reusable, 3D-printed, peroxidase mimic–incorporating multi-well plate for high-throughput glucose determination

[Display omitted] •An incorporation scheme was optimized to functionalize the raw material for 3DP.•The PLA filaments were treated with iron oxide NPs having peroxidase activities.•A 3D-printed 48-well plate efficiently catalyzed the oxidation of TMB by H2O2.•It allowed measuring the absorbance by directly loading it into a plate reader.•The rapid determination of glucose in clinical and food samples was illustrated. Three-dimensional (3D) printing technologies enable rapid laboratory-scale customization of experimental devices to fit the user’s requirements. To demonstrate how 3D-printed objects can be functionalized through incorporation of reactive substances into the raw materials prior to printing, this paper describes a multi-well plate fabricated using polylactic acid (PLA) filaments that had been treated in advance with iron oxide (Fe2O3, Fe3O4) nanoparticles that possess intrinsic peroxidase activities. The fabricated multi-well plate efficiently catalyzed the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) by the peroxidase substrate hydrogen peroxide and allowed measurements of the resulting absorbance through direct loading of the plate into a plate reader. The applicability of this 3D-printed peroxidase mimic–incorporating multi-well plate has been examined in terms of the device’s reusability and analytical performance, analyses of reaction kinetics, and the high-throughput determination of glucose concentrations in clinical and food samples. The results suggest that this functionalization scheme can diversify the utility of current 3D printing technologies in the fabrication of experimental devices with properties comparable with or superior to those of conventional systems.