title: Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk.
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 (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%.
authors.affiliation: Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Huang
authors.forename: Yun
authors.initials: Y
authors.lastname: Yin
authors.forename: Lei
authors.initials: L
authors.lastname: Lu
authors.forename: Hai
authors.initials: H
authors.lastname: Ding
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.forename: Ting
authors.initials: T
authors.lastname: Yu
authors.forename: Jia-jin
authors.initials: JJ
authors.lastname: Zhu
authors.forename: Xiao-dong
authors.initials: XD
authors.lastname: Zheng
authors.forename: Yan-zhen
authors.initials: YZ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002417:Cattle; D002939:Ciprofloxacin / Q000032:analysis* / Q000276:immunology; D003429:Cross Reactions; D004350:Drug Residues / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006241:Haptens / Q000276:immunology; D008297:Male; D008892:Milk / Q000737:chemistry*; D011817:Rabbits
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D006241:Haptens; D002939:Ciprofloxacin