In order to effectively monitor a wide variety of sulfonamides residues in the environment, group-targeting immunoassay based on the group-specific antibodies has attracted great attentions, which can realize the detection of a group of contaminants in environment as many as possible even the unrecognized ones. Indirect competitive immunoassay is generally adopted for small molecule detection however the rational design of immobilized coating antigen for improved recognition capability on the solid surface is far from enough. To cover the research gap, we proposed the design criteria of coating antigen for surface-based indirect competitive immunoassay based on the molecular docking. Taking the group-specific antibodies against sulfonamides (SA) as a proof-of-concept, a hapten with a linking arm with 3 methyl groups was selected to synthesize the coating antigen. Through surface immobilization of coating antigen, a portable biosensor for group-targeting immunoassay of sulfonamides was developed and demonstrated excellent performance with detection limits lower than 0.6 μg/L for four SA variants, and the cross-reactivities of 148-215 % relative to sulfadiazine. The recovery rates of SAs in liquid milk ranges from 87 to 97 %, which confirmed the application potential of this method in the determination of SAs. Its capability to measure total SAs in a simple and low-cost way would pave the way for a variety of application fields.
Keywords: Biosensor; Evanescent wave; Group specificity; Molecular docking; Sulfonamides.
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