ABSTRACT Traditional electroanalytical methods remain a cornerstone in electrochemical biosensor optimizations. In the particular case of chronoamperometry, scalability, cost, and time‐consuming practices challenge to a certain extent its use in multivariate biosensor design workflows. In this study, we demonstrate that spectrophotometric enzymatic assays can be employed as rapid and reliable indicators of the electrochemical response of glucose biosensors, significantly streamlining statistical optimization processes. To this end, second‐generation glucose biosensors were fabricated by co‐depositing flavin adenine dinucleotide‐dependent glucose dehydrogenase (FAD‐GDH) and polydopamine onto graphite electrodes. A two‐stage Design of Experiments (DoE) approach was implemented to systematically evaluate the effects of dopamine and enzyme concentrations. During the exploration step, UV‐Vis measurements of ferricyanide reduction kinetics (ferricyanide being a redox mediator) were used as a proxy for the enzymatic activity, which was compared with the electrochemical sensitivity values obtained via chronoamperometry in the optimization step. A strong correlation between both responses was found, as evidenced by similar predictions of the optimal sensor fabrication conditions, confirming that simple spectrophotometric enzymatic assays can reliably reflect the trends in the biosensor performance. These findings suggest that spectrophotometric measurements offer a rapid, cost‐effective and parallelizable alternative to traditional amperometric workflows, particularly during early stages in multivariate optimizations, implying minimal experimental workload.
Rodriguez et al. (Tue,) studied this question.