Development of a monoclonal antibody-based broad-specificity ELISA for fluoroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds.
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.
Wang
Zhanhui
Z
Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100094, People's Republic of China.
Zhu
Yan
Y
Ding
Shuangyang
S
He
Fangyang
F
Beier
Ross C
RC
Li
Jiancheng
J
Jiang
Haiyang
H
Feng
Caiwei
C
Wan
Yuping
Y
Zhang
Suxia
S
Kai
Zhenpeng
Z
Yang
Xinling
X
Shen
Jianzhong
J
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
D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D024841:Fluoroquinolones; D000077422:Enrofloxacin; D002939:Ciprofloxacin; D015242:Ofloxacin; D009643:Norfloxacin