Preparation of Anti-gatifloxacin Antibody and Development of an Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Gatifloxacin Residue in Milk

Abstract

A polyclonal anti-gatifloxacin antibody has been prepared, and an indirect competitive enzyme-linked immunosorbent assay (cELISA) was developed on the basis of the antibody prepared for the first time. The antibody shows high sensitivity with an IC₅₀ value of 2.6 ppb and excellent specificity with only a minor cross-reaction with lomefloxacin (3.0%) among common (fluoro)quinolones evaluated in this study. The high specificity of the antibody was explained by the molecular structures of related drugs by comparison with published research. The cELISA test kit developed has a detection limit of 0.05 ppb and could be used as a screening method to detect and regulate illegal use of gatifloxacin in food and food products. The test kit was applied to the detection of milk samples spiked by gatifloxacin. The recovery rates were in the range of 86−106%, whereas the intra- and interassay coefficients of variation were <14.3 and <19.6%, respectively. Keywords: Gatifloxacin; drug residue; antibody; immunoassay; cross-reactivity