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CLC number: TS207.3

On-line Access: 2024-08-27

Received: 2023-10-17

Revision Accepted: 2024-05-08

Crosschecked: 2010-09-09

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Journal of Zhejiang University SCIENCE B 2010 Vol.11 No.10 P.812-818

http://doi.org/10.1631/jzus.B1000055


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


Author(s):  Bin Huang, Yun Yin, Lei Lu, Hai Ding, Lin Wang, Ting Yu, Jia-jin Zhu, Xiao-dong Zheng, Yan-zhen Zhang

Affiliation(s):  Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China; more

Corresponding email(s):   zyz85@hotmail.com

Key Words:  Ciprofloxacin, Competitive enzyme-linked immunosorbent assay (ELISA), Rabbit monoclonal antibody, Residues, Cross-reactivity


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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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