Sensitivity-improved nanozyme LFIA with optimized fluid control on centrifugal microfluidic platform

Due to the inflexible manual operation, the sensitivity of the nanozyme-based lateral flow immunoassay (LFIA) can be potentially limited by incontrollable chromatography, insufficient utilization of sample or reagent, and suboptimal binding efficiency. Here, we present a centrifugal microfluidic platform to improve the sensitivity of nanozyme LFIA with multiple strategies to not only automate the entire assay but also to achieve more thorough reaction at each step. Two different pneumatic siphon valves without hydrophilic treatment are developed to facilitate homogeneous mixing and controllable discharging for both sample and chromogenic substrate based on two different deceleration conditions, respectively. In the first step, sustained, smooth and slow sample delivery to the strip is achieved to allow size-controllable sample loading without sacrificing the consistence of chromatograph. In the second step, to increase the reaction time of the chromogenic substrate, the chip is kept centrifuging in chromatograph process to slow down the lateral flow speed for more efficient reaction. Moreover, the chip design is further optimized to reduce the unexpected sample or reagent loss inside the chip. The performance of the developed system is systematically evaluated, and improved detection sensitivity is achieved based on comprehensive optimization, which is beneficial to POC diagnosis.