A Homogeneous Fluorescence Polarization Immunoassay for the Determination of Cephalexin and Cefadroxil in Milk

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• 作者：Jing Zhang (1) (2)
  Zhanhui Wang (1) (2)
  Tiejun Mi (1) (2)
  Lequn Wenren (1) (2)
  Kai Wen (1) (2)
  
• 关键词：Fluorescence polarization immunoassay ; Monoclonal antibody ; Cephalexin ; Cefadroxil ; Milk
• 刊名：Food Analytical Methods
• 出版年：2014
• 出版时间：April 2014
• 年：2014
• 卷：7
• 期：4
• 页码：879-886
• 全文大小：640 KB
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• 作者单位：Jing Zhang (1) (2)
  Zhanhui Wang (1) (2)
  Tiejun Mi (1) (2)
  Lequn Wenren (1) (2)
  Kai Wen (1) (2)
  
  1. College of Veterinary Medicine, China Agricultural University, Beijing, 100093, People’s Republic of China
  2. National Reference Laboratories for Veterinary Drug Residue, District Haidian, Beijing, 100193, People’s Republic of China
  
• ISSN：1936-976X

文摘

A new monoclonal antibody (mAb) against cephalexin (CEX), named 1F3, was produced by immunizing mice with immunogen CEX–keyhole limpet hemocyanin. Two fluorescein-labeled tracers (CEX–fluorescein isothiocyanate (FITC) and cefadroxil (CDX)–FITC) were synthesized and purified by thin-layer chromatography. A homogeneous fluorescence polarization immunoassay (FPIA) based on the new mAb and the selected tracer for the simultaneous determination of CEX and CDX was developed and optimized. The optimized FPIA provided a limit of detection of 1.8 and 2.3?ng?ml? for CEX and CDX and exhibited cross-reactivities of 100 and 63.16?% for CEX and CDX, respectively, and <1?% for other 11 cephalosporins. Recoveries from fortified milk were ranged from 82 to 125?% with coefficient of variations of 3.2-4.2?%. The results indicated that the homogeneous FPIA we developed was enough sensitive to rapidly monitor CEX and CDX in milk.