Preparation of monoclonal antibody and development of an enzyme-linked immunosorbent assay for the detection of ceftiofur in animal-derived foods

Abstract

Ceftiofur (CEF) residues in animal-derived foods are of great concern to farmers, regulatory agencies and consumers. In this study, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method was established to quickly monitor CEF residues in edible animal tissues using an easy sample preparation procedure. A monoclonal antibody, 4D5, against CEF has been produced at first, which had IC50 values for CEF, ceftriaxone, cefquinome, cefotaxime and desfuroylceftiofur of 0.78 μg/L, 0.73 μg/L, 13.6 μg/L, 8.99 μg/L and 8.89 μg/L, respectively. The limit of detection (LOD) and limit of quantitation (LOQ) in artificially contaminated animal-derived foods were 0.12–0.19 μg/L and 0.20–0.30 μg/L. The recovery rates were in the range of 89.7–109.0%. The CVs were less than 6.7%. A good correlation (R= 0.9994) between the ic-ELISA and UPLC-MS/MS showed the reliability of the developed ic-ELISA. The ic-ELISA produces a sensitive, accurate and low-cost tool for the screening of residues of CEF in animal-derived foods.