Abstract
Fast and sensitive screening of antibiotics is a great need in food quality assurance. A monoclonal antibody specific to fluoroquinolones (FQ) was obtained using ciprofloxacin (CPF) derivant as hapten. The antibody was characterized with broad recognition to CPF (100 %), enrofloxacin (ENF, 73.89 %), norfloxacin (73.57 %), nadifloxacin (67.28 %), danofloxacin (53.09 %), pefloxacin (50.26 %), lomefloxacin (35.66 %), enoxacin (12.40 %), and sarafloxacin (3.23 %). Then, an enzyme linked immunosorbent assay (ELISA) was established. The optimized concentrations of coating antigen and antibody were 0.1 and 0.5 μg/ml, respectively. Various parameters including pH values, ionic strength and the concentration of antibody and antigen were optimized for this assay. The ELISA was found to have the best sensitivity in assay buffer of pH 6 and sodium chloride content 1.6 % (m/v) using CPF as tested compound. Fortified milk samples with CPF and ENF (50 and 250 μg/l) and fortified chicken samples (10 and 50 μg/kg) were analyzed with the proposed measure. The ELISA was examined with recovery range of 94–104 % for milk detection and 93–108 % for chicken detection. The coefficient variation data for intra-assay and inter-assay ranged from 4.24 % to 12.16 %. All results indicate the potential extensive application prospects of this ELISA in FQ monitoring for food safety.
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Acknowledgments
This work is financially supported by the Key Programs from MOST (2012AA06A303, 2012BAD29B04).
Conflict of Interest
Steven Suryoprabowo declares that he has no conflict of interest. Liqiang Liu declares that he received grants from MOF and MOE (201310135, JUSRP51308A), which were provided by Jiangsu Province government, China. Juan Peng declares that she has no conflict of interest. Hua Kuang has received national research grants from Key Programs from MOST (2012BAC01B07, 2012AA06A303). Chuanlai Xu has received national research grants from Key Programs from MOST 2012BAD29B04, 2012 BAK11B01). This article does not contain any studies with human subjects. All animal studies were carried out under the guidance of animal welfare committee of Jiangnan University.
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Suryoprabowo, S., Liu, L., Peng, J. et al. Development of a Broad Specific Monoclonal Antibody for Fluoroquinolone Analysis. Food Anal. Methods 7, 2163–2168 (2014). https://doi.org/10.1007/s12161-014-9863-1
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DOI: https://doi.org/10.1007/s12161-014-9863-1