Multifunctional bacteria-derived tags for advancing immunoassay analytical performance with dual-channel switching and antibodies bioactivity sustaining

Constructing multifunctional immunochromatographic assays (ICA) carriers with multiple signals and retaining bioactivity of monoclonal antibodies (mAbs) are conducive to the sensitive and accurate point-of-care testing (POCT). To fulfill this pressing need, a microorganism-based microsphere mediated dual-modal ICA (DICA) was developed for sensitive and reliable detection of zearalenone (ZEN). As the key to the biosensor, a superb biotag with an intact coccus morphology was designed based on Staphylococcus aureus biosynthesized quantum dots incorporating Ru(bpy)(3)(2+) (SAQDsRu), in which SA offered a specific recognition capacity for Fc region of mAbs, QDs endowed a naked-eye discernible colorimetric signal on the SA, and robust fluorescence signal that remedied for the insufficient brightness of QDs was derived from Ru(bpy)(3)(2+). The characterization of SAQDsRu-labeled mAb (SAQDsRu-mAb) probe demonstrated strong luminescence, excellent stability and high affinity with ZEN (affinity constant was approximately 1.723 x 10(9) M-1), which can significantly improve the detection sensitivity. Impressively, a portable sensing system was developed by the integration of SAQDsRu-DICA with a smartphone-based readout. After optimization, this DICA indicated a limit of detection reaching down to 0.008 ng/mL (colorimetric mode) and 0.0058 ng/mL (fluorescent mode), which were much lower than that of conventional gold nanoparticles-based ICA (0.1029 ng/mL), possessing favorable specificity and repeatability (relative standard deviation (RSD) < 10%). Moreover, the feasibility of the immunoassay was further assessed by measuring ZEN in real samples with satisfactory recoveries, and the results are good consistent with liquid chromatography tandem mass spectrometry (LC-MS/MS).