Sandwich-type electrochemical immunosensor based on nitrogen-doped porous carbon and nanoporous trimetallic nanozyme (PdAgCu) for determination of prostate specific antigen

A sandwich-type electrochemical immunosensor was designed for the ultrasensitive detection of prostate-specific antigen (PSA), using Au nanoparticles (Au NPs) modified nitrogen-doped porous carbon (NPC) as sensor platform and trimetallic PdAgCu mesoporous nanospheres (PdAgCu MNSs) as enzyme-mimicking labels. NPC was prepared by a facile one-step pyrolysis strategy of biomimetic phylloid zeolite imidazole framework (ZIF-L) nanosheets. Through this strategy, the graphitization of the microcrystalline structure enhanced the electrical conductivity, while its enlarged specific surface area and abundant pore volume can enrich H₂O₂ to improve the catalytic efficiency. Moreover, Au NPs were used to modify NPC without cross-linking agents to further optimize electron transport while capturing primary antibodies, improving stability and sensitivity of the immunosensor. PdAgCu MNSs with uniform size, cylindrical open mesoporous channels, and continuous crystal frame structure were self-assembling synthesized by electrostatic adsorption and ascorbic acid (AA) co-reduction with amphiphilic dioctadecyldimethylammonium chloride (DODAC) as surfactant-cum-micelle, whose unique structure maximizes the use of polyatoms to expose catalytic sites, exhibiting good biocompatibility and electrocatalytic ability. Under the optimal conditions, the immunosensor showed superior sensitivity, a wide dynamic detection range (10 fg mL⁻¹ ~ 100 ng mL⁻¹) and a low limit of detection (LOD, 3.29 fg mL⁻¹). This work provides a convenient strategy for the clinical detection of PSA. Graphical abstract