Preparation of Broadly Specific Monoclonal Antibodies for Simultaneous Determination of Fluoroquinolone Residues in Eggs
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Preparation of Broadly Specific Monoclonal Antibodies for Simultaneous Determination of Fluoroquinolone Residues in Eggs Dapeng Peng 1 & Yulian Wang 1 & Liang Feng 1 & Guangcai Cao 1 & Yanfei Tao 1 & Zhenli Liu 1 & Zonghui Yuan 1
Received: 20 November 2015 / Accepted: 20 May 2016 # Springer Science+Business Media New York 2016
Abstract To quickly monitor fluoroquinolone (FQ) residues in eggs, three broadly specific monoclonal antibodies (mAbs; i.e., 5B11, 4B6, and 6A2) against FQs were generated. Among them, the 6A2 mAb exhibited the capability to recognize 19 FQs with equal or nearly equal efficacy and therefore was selected for the development of a simple and rapid indirect competitive enzymelinked immunosorbent assay (ic-ELISA). The optimized icELISA protocol was carried out using only acetonitrile with NaOH for the fast extraction of FQs in whole eggs. The CCα, the CCβ, and the limit of quantity values of 19 FQs in eggs ranged from 0.6 to 3.1, 0.7 to 3.6, and 1.6 to 4.2 μg kg−1, respectively. The recoveries from eggs spiked with the above FQs ranged from 60.2 to 119.9 %, with coefficients of variation (CVs) of less than 17.8 %. These results suggest that the prepared mAbs and the developed ic-ELISA protocol are reliable tools that can be applied for the screening of FQ residues in eggs. Keywords Fluoroquinolones . Indirect competitive enzyme-linked immunosorbent assay . Monoclonal antibody . Egg . Residues
Introduction Fluoroquinolones (FQs), which are derived from 3quinolonecarboxylic acid, have a fluorine substituent at the * Zonghui Yuan [email protected] 1
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China
position 6 and are a group of structurally related synthetic antibacterial agents (Spoo and Riviere 1995). These compounds exert their effects by inhibiting DNA gyrase or topoisomerase II in susceptible bacteria and exhibit high activity against a broad spectrum of Gram-negative and Gram-positive bacteria (Bryskier 1993; Drlica 1999). Therefore, these agents are widely used for therapy and prevention of diseases in food-producing animals, aquaculture, pets, and humans. However, the extensive and/or extra-label use of FQs has resulted in the potential presence of residues of these compounds in edible animal tissues or products, which could cause unwanted reactions in humans. In addition, FQ residues have even been detected in a wide range of environments due to the incomplete removal in excreta of humans and animals in wastewater treatment plants and the use of animal excreta as agricultural fertilizers as well as direct discharge from aquaculture treatments, which may pose a threat to various organisms (Zhang et al. 2013). Therefore, the maximum residue limits (MRLs) have been established at different levels for several FQs, including danofloxacin (DAN), enrofloxacin (ENR), flume
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