Journal of Food, Agriculture and Environment

Vol 12, Issue 2,2014
Online ISSN: 1459-0263
Print ISSN: 1459-0255

Development of a monoclonal antibody-based broad-specificity ELISA for fluoroquinolone residues


Jin-qing Jiang

Recieved Date: 2014-01-20, Accepted Date: 2014-03-08


A broad specificity ELISA based on monoclonal antibody for multiresidue determination of fluoroquinolone (FQ) residues has been developed. For this purpose, pefloxacin (PEF) was employed to synthesize the immunogen, and cell fusion technology was used to produce anti-PEF monoclonal antibody. Five hybridoma cell lines named PEF-1, PEF-2, PEF-3, PEF-4 and PEF-5 were screen out, their corresponding mAbs were of the IgG1 isotype with k light chain, and the Kaffs of all mAbs were between 2.2 and 4.5×109 L/mol. Based on the square matrix titration, an icELISA method was established, and its dynamic range for pefloxacin in assay buffer was from 0.008 to 96 ng/mL, with LOD and IC50 value of 0.005 ng/mL and 0.6 ng/mL, respectively. The results of this assay showed a high cross-reactivity to ciprofloxacin (80%), enrofloxacin (70%), difloxacin (68%), sarafloxacin (63%), and norfloxacin (59%). Under 20-fold dilution in authentic milk sample, the matrix effects actually can be ignored. The results suggest this immunoassay can be used for simultaneous detecting six kinds of FQs in milk samples.


Pefloxacin, fluoroquinolones, monoclonal antibody, indirect competitive ELISA, milk

Journal: Journal of Food, Agriculture and Environment
Year: 2014
Volume: 12
Issue: 2
Category: Food and Health
Pages: 70-73

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