title: Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp.
abstract: 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 (IC50) and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an IC50 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.
authors: Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand; Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok 10330, Thailand; Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors: Tossapon
authors: T
authors: Wongtangprasert
authors: Wirongrong
authors: W
authors: Natakuathung
authors: Umaporn
authors: U
authors: Pimpitak
authors: Anumart
authors: A
authors: Buakeaw
authors: Tanapat
authors: T
authors: Palaga
authors: Kittinan
authors: K
authors: Komolpis
authors: Nanthika
authors: N
authors: Khongchareonporn
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002417:Cattle; D004350:Drug Residues / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D020128:Inhibitory Concentration 50; D051379:Mice; D008807:Mice, Inbred BALB C; D010118:Oxytetracycline / Q000032:analysis / Q000737:chemistry*; D033561:Penaeidae / Q000737:chemistry*; D015203:Reproducibility of Results; D012382:Rolitetracycline / Q000737:chemistry; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine
chemical_list: D000911:Antibodies, Monoclonal; D012710:Serum Albumin, Bovine; D012382:Rolitetracycline; D010118:Oxytetracycline