Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

• Published in: Journal of Zhejiang University. B. Science 2014-02, Vol.15 (2), p.165-172
• Main Authors: Wongtangprasert, Tossapon, Natakuathung, Wirongrong, Pimpitak, Umaporn, Buakeaw, Anumart, Palaga, Tanapat, Komolpis, Kittinan, Khongchareonporn, Nanthika
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Biomedical and Life Sciences; Biomedicine; Biotechnology; Cattle; Drug Residues - analysis; Enzyme-Linked Immunosorbent Assay; Female; Inhibitory Concentration 50; Mice; Mice, Inbred BALB C; Oxytetracycline - analysis; Oxytetracycline - chemistry; Penaeidae - chemistry; Reproducibility of Results; Rolitetracycline - chemistry; Sensitivity and Specificity; Serum Albumin, Bovine
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.B1300181

A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2-4F, 7-3G, and 11-11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC 50 ) and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an IC 50 of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross-reactivity to other unrelated antibiotics. When MAb 2-4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%–118% for an intra-assay and 96%–113% for an inter-assay. The coefficients of variation of the assays were 3.9%–13.9% and 5.5%–14.9%, respectively.