Advantages of aggregation-induced luminescence microspheres compared with fluorescent microspheres in immunochromatography assay with sandwich format

Organic fluorescein dye-embedded fluorescent microspheres (FMs) are currently the most established commercially fluorescent markers, and they have been widely used to improve the sensitivity of immunochromatography assay (ICA). However, these FMs have natural defects, such as the aggregation-caused quenching effect and small Stokes shift, which are not conducive to improving the detection performance of ICA. Herein, two green emitted FMs, namely aggregation-induced emission FMs (AIEFMs) and fluorescein isothiocyanate FMs (FITCFMs), were prepared by swelling the AIE luminogens and FITC dyes into the carboxyl group-modified polystyrene microspheres. The average diameters of AIEFMs and FITCFMs were 350 and 450 nm, respectively. Compared with FITCFMs, the AIEFMs exhibited stronger fluorescence intensity and a larger Stokes shift. These two FMs were used as the labeling markers of ICA for procalcitonin (PCT) detection with the sandwich format. Among them, AIEFM-ICA showed dynamic linear detection of PCT from 7.6 pg mL−1 to 125 ng mL−1 with the limit of detection (LOD) at 3.8 pg mL−1. These values were remarkably superior to those of FITCFM-ICA (linear range from 61 pg mL−1 to 62.5 ng mL−1 and LOD value at 60 pg mL−1). Furthermore, the average recoveries of the intra- and inter-assays of AIEFM-ICA ranged from 86% to 112%, with coefficients of variation ranging from 1.2% to 8.8%, indicating accuracy and precision for PCT quantitative detection. Additionally, the reliability of the developed AIEFM-ICA was further assessed by analyzing 30 real serum samples from systemic inflammatory response by infectious diseases, and the results showed good agreement with the chemiluminescence immunoassay. In conclusion, compared with traditional FITCFMs, green emitted AIEFMs as a novel fluorescent label, exhibits greater potential to enhance the detection performance of the ICA platform.