Development of a Monoclonal Antibody-Based Broad-Specificity ELISA for Fluoroquinolone Antibiotics in Foods and Molecular Modeling Studies of Cross-Reactive Compounds

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• 作者：Zhanhui Wang ; Yan Zhu ; Shuangyang Ding ; Fangyang He ; Ross C. Beier ; Jiancheng Li ; Haiyang Jiang ; Caiwei Feng ; Yuping Wan ; Suxia Zhang ; Zhenpeng Kai ; Xinling Yang ; Jianzhong Shen
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：June 15, 2007
• 年：2007
• 卷：79
• 期：12
• 页码：4471 - 4483
• 全文大小：537K
• 年卷期：v.79,no.12(June 15, 2007)
• ISSN：1520-6882

文摘

A competitive indirect enzyme-linked immunosorbentassay (ciELISA) using monoclonal antibodies (Mabs)having broad specificity for fluoroquinolone (FQ) antibiotics is described. Four FQs, ciprofloxacin (CIP), enrofloxacin (ENR), norfloxacin (NOR), and ofloxacin (OFL),were conjugated to bovine serum albumin for immunogens and to ovalbumin for coating antigens. A MabC4A9H1 raised against the CIP hapten exhibited highcross-reactivity (35-100%) with 12 of 14 FQs anddetected these FQs in a ciELISA below their maximumresidue levels (MRLs) with good sensitivity at 50% bindinginhibition (IC₅₀). The quantitative structure-activity relationship (QSAR) between Mab C4A9H1 and variousFQs by comparative molecular field analysis (CoMFA)showed a high predictive ability with a cross-validationq² value of 0.866. Using a simple purification process andthe broad-specificity ciELISA adapted for analysis of FQsin chicken muscle, chicken liver, honey, shrimp, andwhole egg samples demonstrated recoveries of 60-93%for CIP, ENR, NOR, OFL, flumequine, and danofloxacin.