Hand-Operated, Paper-Based Rotational Vertical-Flow Immunosensor for the Impedimetric Detection of alpha-Fetoprotein

This study demonstrates a hand-operated, paper-based rotational vertical-flow immunosensor (rotational VFI) platform requiring fewer pipetting steps, designed for the electrochemical detection of alpha-fetoprotein with multiple and time-sequenced steps. The platform allows users to perform electrochemical measurements without interference from the convective component of fluid motion, which is unfavorable in most techniques. Users can freely transfer-switch-stop fluid flows by manually rotating the paper disk, evidencing the superior flexibility of this sensor compared to other biosensors. Furthermore, the overall assay duration can be considerably shortened to 9 min. The linear range (LR) is determined to be 0.01-500 ng/mL, with a limit of detection (LOD) of 1.65 pg/mL, and the sensitivity can be significantly enhanced simply by switching off the sample stream to ensure detention at the binding zone (for up to 30 min). This additional step can widen the LR to 0.5 pg/mL, with a LOD of 3.54 fg/mL, which is the lowest detectable level ever reported among paper-based sensors. The advantages of the designed rotational VFI qualify it as a suitable alternative to various biosensors.