Controlling the amount of coupling agents on the synthesis of coating antigens to enhance the sensitivity of fluoroquinolone immunodetection

There is now increasing demand to improve the sensitivity of various immunoassays for fluoroquinolones (FQs) and other food hazards. In this study, different coating antigens were prepared by adjusting the content of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) to explore its influence on the immunoassay sensitivity of FQs. The results indicated that, unlike traditional assumptions, a reasonable EDC dosage should be addressed to reach the best analytical efficiency, and excessive EDC could enhance the hapten-carrier conjugation but significantly reduce the detection sensitivity. For the FQs investigated, the hapten:EDC:BSA proportion of 20:2.5:50 (Mole ratio:74:34:1) seemed the best for preparation of coating antigens, and the sensitivity could be improved more than 1000 times both for indirect competitive enzyme linked immunosorbent assay ELISA (ic-ELISA) and gold immunochromatography assay (GICA) due to two key factors including coupling-ratios and amide bond groups. Such an improved efficiency was also validated well with different food samples, which indicated the reasonable optimization of EDC in coating antigen synthesis may be widely used as a new, simple and more effective strategy to improve the immunoassay for low molecular targets in medical, environment and food detection filed.