Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

作者：Bin Huang, Yun Yin, Lei Lü, Hai Ding, Lin Wang, Ting Yu, Jia-jin Zhu, Xiaodong Zheng, Yanzhen Zhang · 发表于：Journal of Zhejiang University SCIENCE B · 年份：2010 · DOI：10.1631/jzus.b1000055 · 被引用次数：47 · 研究领域：Protein purification and stability、Monoclonal and Polyclonal Antibodies Research、Escherichia coli research studies

A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC(50)) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%-84.60%, with coefficients of variation of less than 12.2%.