Detection of enrofloxacin by flow injection chemiluminescence immunoassay based on cobalt hydroxide nanozyme

The emergence and development of low-cost and high-efficiency nanozymes are promising to replace natural enzymes promoting the application of chemiluminescence immunoassays. Herein, a rapid and highly sensitive flow injection chemiluminescence immunoassay based on cobalt hydroxide (Co(OH)₂) nanozyme was established to detect enrofloxacin (ENR) residues in food. In this system, Co(OH)₂ nanosheets act as nanozymes to catalyze and amplify the chemiluminescence signal of the luminol-PIP-H₂O₂ system, as well as a carrier for immobilizing antibodies to form stable immunoprobes. In addition, carboxyl resin beads with good stability and biocompatibility were used as the base of the immunosensor to carry more coating antigens, based on the principle of competitive immunity and to achieve the rapid detection of ENR. Under optimal conditions, the linear working range is 0.0001 ~ 1000 ng/mL, and the limit of detection (LOD) is 0.041 pg/mL (S/N = 3). The method has been successfully applied to the analysis of aquatic products and poultry food. A non-enzyme immunosensor using Co(OH)₂ nanosheets as antibody-conjugated carriers and peroxidase mimics for catalytic amplification of the chemiluminescence signal of luminol and using carboxyl resin beads as platform was designed to detect ENR residues in food.