Immunochromatographic assay based on high-affine monoclonal antibody for the detection of imidocarb in milk.

Abstract

Imidocarb (IM) is an antiprotozoal agent, which is mainly used to treat babesiosis and piroplasmosis for animals. However, overdose or improbable utilization cause IM residues in animal origin products, which would be harmful to human health. Here, a monoclonal antibody (mAb) against IM with extremely sensitive and specific features has been successfully prepared from a novel immunogen synthesized by virtue of the active ester method. The concentration of half-maximal inhibition (IC50 ) of the mAb was 0.074ng/ml and the affinity constant was 4.58 × 1010 L/mol. On the basis of this condition, an immunochromatographic strip (ICS) is proposed that could be applied in milk samples to test IM rapidly. For the ICS, the visual detection limit (cut-off value) was 5ng/ml, IC50 was 0.4ng/ml, the limit of detection (LOD) was 0.078ng/ml, the linear detection scope was 0.117 to 1.37ng/ml. The recovery rates ranged from 88.83% to 91.47% and coefficients of variation (CV) were in the spectrum of 7.31% to 9.43%. In general, the recommended test strip provided an exceedingly simple and reliable detection method for quickly testing the IM. PRACTICAL APPLICATION: In our joint efforts, an extremely sensitive monoclonal antibody against imidocarb was obtained and a test strip for the rapid detection of imidocarb in milk was developed. The developed method could be applied to the field and provided great potential for analytical of imidocarb in other matrixes.