Homogeneous time-resolved fluoroimmunoassay of microcystin-LR using layered WS2nanosheets as a transducer

A homogeneous time-resolved fluoroimmunoassay method for rapid and sensitive detection of microcystin-LR (MC-LR) in water samples was developed based on the interaction between water-soluble WS₂ nanosheets and the conjugate of MC-LR with a luminescent Eu³⁺ complex BHHBCB-Eu³⁺ (BHHBCB: 1,2-bis[4'-(1″,1″,1″,2″,2″,3″,3″-heptafluoro-4″,6″-hexanedion-6″-yl)- benzyl]-4-chlorosulfobenzene). The large lateral dimensions and high surface areas of two-dimensional layered WS₂ nanosheets enable easy adsorption of the MC-LR-BHHBCB-Eu³⁺ conjugate, that lead to efficient quenching of the luminescence of Eu³⁺ complex via energy transfer or electron transfer process. However, the addition of monoclonal anti-MC-LR antibody can induce the formation of MC-LR-BHHBCB-Eu³⁺/antibody immune complex, which prevents the interaction between WS₂ nanosheets and MC-LR-BHHBCB-Eu³⁺ to result in the restoration of Eu³⁺ luminescence. This signal transduction mechanism made it possible for analysis of the target MC-LR in a homogeneous system. The present method has advantages of rapidity and simplicity since the B/F (bound reagent/free reagent) separation steps, the solid-phase carrier and antibody labeling or modification process are not necessary. The proposed immunosensing system displayed a wide linear range, good precision and accuracy, and comparable sensitivity with a detection limit of 0.3 μg l^-1, which satisfied the World Health Organization (WHO) provisional guideline limit of 1.0 μg l^-1 for MC-LR in drinking water.