Abstract
Nanomaterials with intrinsic peroxidase-like activities have been demonstrated as promising artificial enzymes for numerous biochemical applications. In recent years, three dimensional printing (3DP) technologies have accelerated the customization and rapid prototyping of multilayer components/devices in general laboratories for a wide range of analytical applications. In this study, to enable the 3D-printed devices to have both the peroxidase and chromogenic activities, a multi-well plate is one-step printed using a dual-head fused deposition modeling-type 3D printer with two functionalized thermoplastic filaments, acrylonitrile butadiene styrene incorporated with peroxidase-mimicking iron oxide (Fe3O4) nanoparticles and polyvinyl alcohol infiltrated with chromogenic substrate o-phenylenediamine (OPD). The fabricated plates are confirmed to efficiently catalyze the oxidation of OPD by peroxidase substrate H2O2 and allow measurement of the absorbance of the derivatized samples through directly loading it into a conventional plate reader. After method’s optimization, the method’s detection limits can reach as low as 2.8 μM for H2O2 and 5.0 μM for glucose. Based on our demonstrations, the proposed 3D-printed peroxidase mimic/chromogenic substrate-incorporated multi-well plates can provide the competitive peroxidase activities, in-well derivatization and measurement/visualization without any sample transfer, and the applicability of in situ, rapid, high-throughput analyses of glucose concentrations in clinical samples.