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Abstract. Antibiotic residues in animal-derived food products have been identified as a potential hazard in human health. Hence, a rapid, simple and cost-effective method for antibiotic detection in food supply chain is highly desirable. The objective of this study was to develop a nanomaterials-based biosensor using immunomagnetic beads (IMBs) and quantum dots (QDs) for rapid and sensitive detection of enrofloxacin (ENR) in chicken products. A 5-sulfosalicylic acid-based pretreatment method was adopted to reduce non-specific adsorption caused by complex food matrices. Two sensing elements, antibody functionalized magnetic beads and enrofloxacin-bovine serum albumin conjugates modified QDs (QDs-BSA-ENR) were designed and fabricated. Target ENR in samples was first captured and separated by IMBs. Then, QDs-BSA-ENR, served as a competitor and detection probe, was used to react with the residual binding sites on IMBs surface. With the presence of captured ENR, the binding of QDs-BSA-ENR to IMBs surface was competitively inhibited. Finally, the fluorescence of all reporting QDs in the QDs-BSA-ENR-IMBs complex was measured for the quantitation of target antibiotics. Under the optimum conditions, the proposed method allowed for sensitive detection of ENR in a linear range from 1 to 100 ng mL^-1 with a limit of detection (LOD) of 0.94 ng mL^-1. The LOD for spiked chicken muscle samples was 16.26 μg kg^-1, which was below the maximum residue limits regulated in China and the European Union. The whole analytical procedure from food sampling to result report could be finished within 1 h. This nanobiosensor showed high potential for rapid, low-cost and in-field detection of ENR in poultry supply chain to enhance food safety.