Nanozyme amplification mediated on-demand multiplex lateral flow immunoassay with dual-readout and broadened detection range

Development of sensitive, facile and rapid biosensors is important for widespread applications. Nanozymes can be ideal signal donors for constructing dual-readout lateral flow immunoassays (LFIA) because they are an excellent class of optical reporters. Herein, a magnetic prussian blue nanozyme (MPBN) mediated dual-readout on-demand multiplex lateral flow immunoassay (MLFIA) was established by employing ractopamine (RAC) and clenbuterol (CLE) as the model analytes. The MPBN was synthesized through in-suit shell-growing and introduced as a bifunctional signal tag owing to their darker original color and peroxidase-like activity. Based on the catalytic signal created by catalyzing oxidation of chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) and colorimetric signal generated by tag's original color, improved precision and broadened detection range were acquired by implementing a dual-readout strategy. And a two-fold increase in the detection range could fulfill different limit requirements of the same target in various regions. The obtained recoveries from 84.01% to 119.94% indicating the repeatability and reliability of the proposed method. This method provides an attractive platform for the detection of a same target with different detection limits, which possesses a considerable potential in monitoring of other targets.