摘要Herein,a novel label-free electrochemical immunosensor was fabricated via immobilizing specific anti-β-lactoglobulin(β-LG)antibodies(Abs)onto an integrated electrode of gold nanoparticles(AuNPs)/Prussian blue(PB)/cubic Ia3d structured mesoporous carbon(CMK-8).This immunosensor allowed for the quantitative detection of the major milk allergen β-LG.CMK-8 with excellent electrical conductivity and uniformly adjustable pore structure was modified on the glassy carbon electrode(GCE)and served as the sensitive substrate for the electro-polymerization of PB,forming the redox-active layer.AuNPs were subsequently electrochemically deposited on PB/CMK-8/GCE to improve the electrical conductivity and utilized as the connector for Abs immobilization.During β-LG detection,the Abs-modified AuNPs/PB/CMK-8/GCE exhibited a significant reduction in differential pulse voltammetry current signal when exposed to β-LG,displaying an inverse dose-dependent relationship.The developed electrochemical immunosensor demonstrated good detection performance for β-LG,with a wider linear range of 0.01-100 ng/mL and a lower detection limit of 4.72 pg/mL.Meanwhile,the sensor exhibited remarkable repeatability,reproducibility,stability and anti-interference capabilities,which was further applied to detect β-LG in dairy food,achieving satisfactory recoveries(89.2%-98.8%)and lower relative standard deviation(￡ 3.1%).Therefore,this innovative electrochemical method for food allergen detection holds great potential application in food safety determination and evaluation.