title: Development of a monoclonal antibody-based broad-specificity ELISA for fluoroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds.
abstract: A competitive indirect enzyme-linked immunosorbent assay (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 Mab C4A9H1 raised against the CIP hapten exhibited high cross-reactivity (35-100%) with 12 of 14 FQs and detected these FQs in a ciELISA below their maximum residue levels (MRLs) with good sensitivity at 50% binding inhibition (IC50). The quantitative structure-activity relationship (QSAR) between Mab C4A9H1 and various FQs by comparative molecular field analysis (CoMFA) showed a high predictive ability with a cross-validation q2 value of 0.866. Using a simple purification process and the broad-specificity ciELISA adapted for analysis of FQs in chicken muscle, chicken liver, honey, shrimp, and whole egg samples demonstrated recoveries of 60-93% for CIP, ENR, NOR, OFL, flumequine, and danofloxacin.
authors.affiliation: Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100094, People's Republic of China.
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhu
authors.forename: Shuangyang
authors.initials: S
authors.lastname: Ding
authors.forename: Fangyang
authors.initials: F
authors.lastname: He
authors.forename: Ross C
authors.initials: RC
authors.lastname: Beier
authors.forename: Jiancheng
authors.initials: J
authors.lastname: Li
authors.forename: Haiyang
authors.initials: H
authors.lastname: Jiang
authors.forename: Caiwei
authors.initials: C
authors.lastname: Feng
authors.forename: Yuping
authors.initials: Y
authors.lastname: Wan
authors.forename: Suxia
authors.initials: S
authors.lastname: Zhang
authors.forename: Zhenpeng
authors.initials: Z
authors.lastname: Kai
authors.forename: Xinling
authors.initials: X
authors.lastname: Yang
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002939:Ciprofloxacin / Q000032:analysis; D000077422:Enrofloxacin; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D024841:Fluoroquinolones / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis*; D020128:Inhibitory Concentration 50; D009643:Norfloxacin / Q000032:analysis; D015242:Ofloxacin / Q000032:analysis; D021281:Quantitative Structure-Activity Relationship; D012680:Sensitivity and Specificity
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D024841:Fluoroquinolones; D000077422:Enrofloxacin; D002939:Ciprofloxacin; D015242:Ofloxacin; D009643:Norfloxacin