Generic Immunoassay of Quinolones: Production and Characterization of Anti-pefloxacin Antibodies as Broad Selective Rece
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Generic Immunoassay of Quinolones: Production and Characterization of Anti-pefloxacin Antibodies as Broad Selective Receptors Limin Cao & Jianxin Sui & Dexin Kong & Zongyan Li & Hong Lin
Received: 6 July 2010 / Accepted: 5 January 2011 / Published online: 21 January 2011 # Springer Science+Business Media, LLC 2011
Abstract Polyclonal antibodies were raised against pefloxacin, and their specificity to different quinolones was investigated and validated. The results of the competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) demonstrated that the mice anti-pefloxacin antibody could efficiently bind with at least nine quinolones, and to six of them, the cross-reactivity was calculated higher than 65%. Exploiting these antibodies as receptors, six quinolones in fortified eel samples were effectively detected by both ciELISA and dot immunogold filtration assay, in which the least detection limit was estimated as ≤10 μg/kg and ≤20 μg/kg, respectively. These results fit well with molecular modeling studies based on field-overlapping and allow us to suggest possible applications of anti-pefloxacin antibody as a new broad selective receptor for generic assay of quinolones. Keywords Quinolone . Broad selective receptor . Pefloxacin . Antibody
L. Cao : J. Sui : Z. Li : H. Lin (*) Food Safety Laboratory, Ocean University of China, Qingdao 266003, People’s Republic of China e-mail: [email protected] D. Kong State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, People’s Republic of China D. Kong Shandong Provincial Research Center for Bioinformatics Engineering and Technique, Center for Advanced Study, Shandong University of Technology, Zibo 255049, People’s Republic of China
Introduction Quinolones are a group of broad-spectrum synthetic antibiotics. Since the 1960s, more than 20 quinolones have been applied for the treatment of bacterial infections in human and animals (Bhanot et al. 2001; Zhou and Zhou 2005). Due to increasing concerns on food safety, many countries and regions have set strict maximum residue limits (MRLs) for quinolone residues in animal products (Hernandez-Arteseros et al. 2002; W. X. Lin (Editor) et al. 2002; Ministry of Agriculture of People’s Republic of China and Agricultural Industry Criteria of People’s Republic of China 2001, 2005). Usually, these regulations are mainly focused on those compounds permitted to be used in veterinary medicines like enrofloxacin, sarafloxacin, and flumequine (Hernandez-Arteseros et al. 2002). Moreover, some other quinolones, often applied illegally in order to charge a lower price or for better antimicrobial efficiency, were also included in the list of drug residues to be monitored (W. X. Lin (Editor) et al. 2002; Ministry of Agriculture of People’s Republic of China and Agricultural Industry Criteria of People’s Republic of China 2001, 2005). Therefore, it becomes an important task now to determine, simultaneously, as many quinolone residues as possible in food samples.
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