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Tossapon Wongtangprasert Wirongrong Natakuathung Umaporn Pimpitak Anumart Buakeaw Tanapat Palaga Kittinan Komolpis Nanthika Khongchareonporn 《Journal of Zhejiang University. Science. B》2014,15(2):165-172
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. 相似文献
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Tossapon WONGTANGPRASERT Wirongrong NATAKUATHUNG Umaporn PIMPITAK Anumart BUAKEAW Tanapat PALAGA Kittinan KOMOLPIS Nanthika KHONGCHAREONPORN 《Journal of Zhejiang University. Science. B》2014,(2)
研究目的:抗土霉素单克隆抗体的制备和特性分析,并用于间接竞争酶联免疫吸附测定法(icELISA)分析虾样品中土霉素的残留。创新要点:本研究建立分泌抗土霉素单克隆抗体的杂交瘤细胞株,筛选出对虾中土霉素残留检测具有较高灵敏度的单克隆抗体2-4F。研究方法:用细胞杂交技术使土霉素-牛血清白蛋白偶联物免疫的BALC/c雌性小鼠的脾细胞与骨髓瘤细胞融合,建立三株分泌抗土霉素单克隆抗体的杂交瘤细胞株(2-4F、7-3G和11-11A),并制备它们的单克隆抗体。通过icELISA法分析单克隆抗体对土霉素的半抑制质量浓度(IC50)和交叉反应率,明确其灵敏度和特异性。重要结论:实验结果发现单克隆抗体2-4F具有最高的灵敏度,对土霉素的IC50为7.01 ng/ml,且该单抗特异性强,仅与罗利环素之间有强烈的交叉反应,而与非相关抗生素不发生交叉反应。当利用单克隆抗体2-4F检测虾样品中的土霉素含量时,其批内和批间回收率分别为82%~118%和96%~113%,变异系数分别为3.9%~13.9%和5.5%~14.9%。因此,单克隆抗体2-4F可用于虾产品中土霉素残留分析试剂盒的开发。 相似文献
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