标题：Preparation of Anti-Lomefloxacin Antibody and Development of an Indirect Competitive Enzyme-Linked Immunosorbent Assay for Detection of Lomefloxacin Residue in Milk
作者：Cao, Zhaozhen; Lu, Shengxin; Liu, Jinting; Zhan, Jinhua; Meng, Meng; Xi, Rimo
作者机构：[Meng, Meng; Xi, Rimo] Nankai Univ, Coll Pharm, Tianjin 300071, Peoples R China.; [Meng, Meng; Xi, Rimo] Nankai Univ, Tianjin Key Lab Mol Drug Res, 更多
通讯作者地址：[Xi, RM]Nankai Univ, Coll Pharm, Tianjin 300071, Peoples R China.
关键词：Antibody; Cross-reactivity; Drug residue; Immunoassay; Lomefloxacin
摘要：Lomefloxacin has been increasingly used in veterinary medicine to treat microbial infections. To avoid using a complicated instrumental method to detect lomefloxacin residue in food, a simple and convenient indirect competitive enzyme-linked immunosorbent assay method has been developed in this study. The antibody generated from immunogen of bovine serum albumin-lomefloxacin showed high sensitivity toward lomefloxacin with an IC50 value of 0.35ppb in PBS buffer and was suitable to be used as a screening assay to detect lomefloxacin residue in food products. The ELISA (Enzyme-Linked ImmunoSorbant Assay) developed in this study was compared with a commercial ELISA kit and significant improvement was achieved in terms of sensitivity and specificity. The antibody prepared showed excellent specificity with certain cross-reactivity with only two drugs including norfloxacin (17.5%) and fleroxacin (8.8%) among commonly used (fluoro)quinolones. The assay measured drug residue in defatted milk spiked with lomefloxacin with an inter-assay coefficient of variation 22.2% and an intra-assay coefficient of variation 18.2%. The average recovery rates at 0.1, 0.3, 0.6, 1.0, and 2.0 ppb were in the range of 120-85% for inter-assay and in the range of 113-90% for intra-assay.