Substitution of antibody with molecularly imprinted 96-well plate in chemiluminescence enzyme immunoassay for the determination of chloramphenicol residues

Abstract

A direct competitive chemiluminescence enzyme immunoassay was developed by using the molecularly imprinted 96-well plate as an artificial antibody to detect chloramphenicol (CAP). The artificial antibody was synthesised on the well surface of MaxiSorp polystyrene 96-well plate and CAP was conjugated with the horseradish peroxidase. The results showed that the imprinted plate exhibited antibody-like binding ability. The plate showed fast adsorption rate, 66% adsorption was finished within 20 min. The cross-reactivity for CAP, florfenicol and thiamphenicol were 100%, 1.25% and 2.08%, respectively. And the imprinted plate could be reused for many times without loss of sensitivity. The IC50 and the detection limit values under optimum conditions were 30±2 µg·L−1 and 0.9±0.01 µg·L−1, respectively. The plate was used to detect CAP in sea cucumber, which showed excellent recoveries ranging from 89% to 98.7%. And the result correlated well with that obtained by the CAP enzyme-linked immunosorbent assay (ELISA) kit.