Anti-oxytetracycline monoclonal antibody based detection of oxytetracycline residue in honey

作者：Xu, Shufa; Hou, Liwei; Wubie, Abebe J.; Cao, Tan; Hu, Yingying; Li, Wei; Guo, Zhanbao; Zhao, Jing^*; Wu, Jie

来源：Journal of Food Agriculture and Environment, 2013, 11(2): 249-257.

• Antibiotics
• antibiotic residue
• oxytetracycline (OTC)
• residue detection
• monoclonal antibody (MAb)
• immunogen
• enzyme-linked immunosorbent assay (ELISA)
• colloidal gold-based immunochromatographic assay (CGIA)

Oxytetracycline (OTC) has the potential to bring severe side effects to humans, when taken from its residues. The enzyme-linked immunosorbent assay (ELISA) technique has long been known as a rapid, sensitive, specific and cost-effective analytical method, and has been used for diagnostic and residue detection purposes for many years. However, there are no reports concerning the synthesis of OTC immunogen and preparation of anti-OTC antibody use in detection of tetracycline family residues in honey so far. Thus, to identify and understand oxytetracycline pollution in beekeeping industry, we prepared an oxytetracycline immunogen and anti-oxytetracycline monoclonal antibody for the detection of its residues in honey. Conjugates of oxytetracycline (OTC); toluidine BAS and toluidine OVA were prepared as an immunogen and has been used to produce monoclonal antibodies (MAb). A hybridoma 3E3 secreting anti-OTC MAb was obtained and has showed a 50% maximal inhibitory concentration (IC50) value of 0.3 ng ml(-1) in phosphate buffered saline. With optimum conditions, an indirect competitive ELISA was developed and applied to detect OTC residues in honey samples. As the result, developed ELISA has shown that minimum detection limit of OTC in honey samples was 1.43 ng g(-1) without cross reactivity to the other tested antibiotics. Optimum CGIA test has also found that minimum detection limit for OTC residues in honey samples to be 2.16 ng g(-1). Furthermore, mean recovery percentage of OTC was found to vary from 83.1% to 102.6%. Furthermore, our work has also verified the efficiency of the technique in the detection of the residues in honey. With these all, results have strongly confirmed and recommended that indirect competitive ELISA and CGIA could be used in screening of OTC residues in honey.

• 出版日期2013
• 单位中国农业科学院