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| Study on a Monoclonal Antibody-based Immunoassay Method for Florfenicol in Animal Meat and Feed |
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| DOI： |
| KeyWord:florfenicol animal meat and feed monoclonal antibody immunoassay |
| Author | Institution |
| XIE Mei-chan，YANG Jin-yi，LI Ran，XIAO Zhili，WANG Hong，XU Zhen-lin，SUN Yuan-ming，SHEN Yu-dong |
Guangdong Provincial Key Laboratory of Food Quality and Safety， College of Food Science， South China Agricultural University， Guangzhou ， China |
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| Abstract: |
| To monitor florfenicol residues in animal-derived foods and feeds， a monoclonal antibody with excellent specificity and sensitivity was prepared by using the antigen reported previously to immunize BALB/c mice and hybridoma technique. An indirect competitive enzyme-linked immunosorbent assay was established based on this antibody. The optimal conditions were finalized as follows： a coating antigen concentration of 0.05 μg/mL， an antibody concentration of 0.1 μg/mL， a competition reaction time of 30 min， a PBST diluent for analyte， primary antibody and secondary antibody with a Tween－20 of 0.05%， a secondary antibody concentration of 0.167 μg/mL and its reaction time of 30 min. Under the optimal conditions， the icELISA method for detection of florfenicol was established with an IC50 of 9.48 ng/mL， a linear range of 1.75－51.36 ng/mL and a limit of detection（LOD）of 0.64 ng/mL. There was no obvious cross-reactivity between the icELISA and its structural and functional analogs such as chloramphenicol， thiamphenicol， etc. The spiked recoveries for florfenicol in samples ranged from 88.1% to 107% with relative standard deviations（RSDs）less than 15%. The developed icELISA method was used to detect fifteen blind samples， of which four blind samples were found to contain florfenicol in different concentrations levels. These results were in good agreement with those of HPLC－MS/MS. Therefore， the developed icELISA exhibited a potential practicality in detection of florfenicol in animal meat and feed samples. |
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