Radical-mediated Electrical Enzyme Assay For At-home Clinical Test

Abstract: To meet the growing demand for accurate, rapid, and cost-effective at-home clinical testing, we developed a radical-mediated enzyme assay (REEA) integrated with a paper fluidic system and electrically read by a handheld field-effect transistor (FET) device. The REEA utilizes horseradish peroxidase (HRP) to catalyze the conversion of aromatic substrates into radical forms, producing protons detected by an ion-sensitive FET for biomarker quantification. Through screening 14 phenolic compounds, halogenated phenols emerged as optimal substrates for the REEA. Encased in an affordable cartridge ($0.55 per test), the system achieved a detection limit of 146 fg/mL for estradiol (E2), with a coefficient of variation (CV) below 9.2% in E2-spiked samples and an r2 of 0.963 across a measuring range of 19 to 4,551 pg/mL in clinical plasma samples, providing results in under 10 minutes. This adaptable system not only promises to offer a fast and reliable platform, but also holds significant potential for expansion to a wide array of biomarkers, paving the way for broader clinical and home-based applications.