Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk |
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| Authors: | Bin Huang Yun Yin Lei Lu Hai Ding Lin Wang Ting Yu Jia-jin Zhu Xiao-dong Zheng and Yan-zhen Zhang |
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| Institution: | 1.Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China;2.Department of Stomatology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China;3.Yikang Biotechnology Company, Hangzhou 310029, China |
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| Abstract: | 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 (IC50) 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%. |
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| Keywords: | Ciprofloxacin Competitive enzyme-linked immunosorbent assay (ELISA) Rabbit monoclonal antibody Residues Cross-reactivity |
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