Fluorescent artificial enzyme-linked immunoassay system based on Pd/C nanocatalyst and fluorescent chemodosimeter

Artificial enzyme mimics have recently attracted considerable interest because they possess many advantages compared with natural enzymes, such as low cost of preparation and high stability. Herein, we present a novel fluorescent artificial enzyme-linked immunoassay strategy by utilizing Pd/C nanocatalyst as the enzyme mimic and bis-allyloxycarbonyl rhodamine 110 (BI-Rho 110) as the substrate, and the amplification procedure is based on the palladium-catalyzed Tsuji-Trost reaction. Pd/C nanocatalyst with the average size of 150 nm was prepared by the impregnation-reduction method, and high resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) analyses reveal that Pd clusters with an average size of about 1 nm are dispersed uniformly on each carbon nanosphere's surface. Kinetic studies show that this reaction follows Michaelis-Menten kinetics and the fluorescence intensity is proportional to the concentration of Pd/C nanocatalyst under certain conditions. The turnover number of Pd/C nanocatalyst reaches up to 3.3 × 10(7) (h(-1)). The analytical performance of this system in detecting hCG shows that after a 24 h incubation the sensitivity limit can reach 0.1 ng/mL and the dynamic linear working range is 1-10 ng/mL. Our findings pave the way to use Pd-catalyzed reaction for design and development of novel analytical methods.