The development of a wash-free homogeneous immunoassay method for the detection of tetracycline in environmental samples

Abstract

Antibiotic residues have become the major source of environmental pollutants. In order to monitor tetracycline (TC) in the environment, we have established a highly sensitive and wash-free homogeneous time-resolved immunoassay. This analytical method was based on a rare earth chelate with excellent fluorescence properties. The cryptate organic ligand had good stability and acted as an antenna for Eu³⁺ excitation. In a homogeneous system, the Eu³⁺ cryptate complex was used as a label to bind to antibodies. Under the action of immunoaffinity, fluorescent donors and acceptors were close to each other, which induced the FRET effect to produce proportional fluorescence. Under the optimal parameters, the half-inhibitory concentration (IC50) and limit of detection (LOD, IC₁₀) of TC were 0.4188 ng mL⁻¹ and 0.0106 ng mL⁻¹, respectively. The linear range (IC20–IC80) was 0.0273–9.2645 ng mL⁻¹. With the environmental samples, the recovery rate of TC was 84.3–107.2%, and the standard deviation (RSD) was 4.6–12.9%. The results showed the good sensitivity and reliability of the method. Compared with the traditional ELISA, our method has less background interference, only one step was required without the washing procedure, and the detection result can be obtained by 30 min incubation, which improves the detection efficiency. Because of the characteristics of immunoassays, different pollutants can be monitored by changing the antibodies. This method provides an alternative path for detecting environmental pollutants and has the potential to develop into an on-site detection kit.