Bionic Enzyme-Assisted Ion-Selective Amperometric Biosensor Based on 3D Porous Conductive Matrix for Point-of-Care Nitrite Testing

Nitrite plays a critical role in a variety of physiological processes and maintaining the nitrite level in an appropriate range is vital to keep healthy. Current nitrite analysis methods lack sensitivity and require tedious operations, which could not meet the need of point-of-care (POC) nitrite detection in precision medicine. Here we present a cyanocobalamin (VB12) bionic enzyme-assisted ion-selective amperometric biosensor based on 3D porous conductive matrix (PCM), which can facilitate rapid and accurate POC nitrite monitoring in complex biofluids. The experimental findings quantitatively demonstrate that the biosensor has a sensitivity of 64.08 μA/(mM·cm2), a wide linear range of 0.025–45 mM, and low limit of detection of 1 nM. Moreover, the developed VB12/BSA-PCM biosensor shows outstanding stability after 21 days with 2% decline in current signal, and high repeatability between batches with RSD of only 1.29%. Real salivary nitrite detection has been evaluated, and the results match well with the commercial nitrite analyzer. Thus, the bionic enzyme-assisted ion-selective amperometric biosensor proposed herein has potential utility as an affordable tool for POC detection and home-based healthcare.