Development of biomimetic enzyme-linked immunosorbent assay based on molecular imprinting technique for semicarbazide detection

Abstract

Herein, we present a direct competitive biomimetic enzyme-linked immunosorbent assay (BELISA) method for the determination of semicarbazide (SEM) using a molecularly imprinted polymer (MIP) film as the bionic antibody. The best monomer (methacrylic acid) and molar ratio of monomer to template (4:1) were determined with the aid of computational simulation methods based on density functional theory (DFT). The MIP film was characterised by scanning electron microscopy and adsorption experiments. The results showed that the imprinted film was successfully synthesised with good adsorption performance. Under optimal conditions, the calibration curve was linear in the range of 1.0–1.0×104 ng/mL (R2 = 0.9863). The detection limit (IC15) and sensitivity (IC50) were 1.0 and 85.7 ng/mL, respectively. This BELISA method was further applied to the analysis of SEM in food samples; recoveries ranged from 86.1% to 90.8%. These observations indicated that the developed method could be adapted for SEM quantification in food.

Keywords

• biomimetic enzyme-linked immunosorbent assay
• molecularly imprinted polymer
• computational simulation
• semicarbazide