Efficient green synthesis of biocompatible MPN fluorescent microspheres via hydrophobic-force-driven strategy for enhanced immunochromatographic assays

The unique fluorescence properties of aggregation-induced emission (AIE) fluorescent microspheres (FMs) make them ideal signal markers. Traditional synthesis methods are complex, labor-intensive, and hazardous, leading to AIEFMs that lack biocompatibility and require further modification for immunoprobe preparation. This study introduces a novel hydrophobic force-driven method for rapid synthesis of highly biocompatible FMs (H-FMs), demonstrating their benefits in immunochromatographic assay (ICA) applications. The metal-polyphenol network (MPN) shell around the AIEgen core structure of H-FMs is quickly and safely formed by depositing MPN onto AIEgen nano-aggregates, achieving high dye utilization, affordability, and design flexibility, while producing H-FMs with fluorescence across 300-800 nm. The excellent biocompatibility of H-FMs eliminates the need for additional modifications, allowing antibodies to be coupled swiftly (within 10 min) with a high coupling efficiency of 93.4 %. The resulting immunoprobes exhibit strong target recognition and 90.6 % fluorescence retention over 30 days. These features support their application in double antibody sandwich and competitive ICA formats, with detection limits of 9.62 × 10² CFU/mL for E. coli O157:H7 and 0.0081 ng/mL for AFM1. This study provides new insights into designing fluorescent probes for safety monitoring of hazardous materials in the environment.