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Fragment-based hapten design and screening of a highly sensitive and specific monoclonal antibody for ractopamine

机译:基于片段的半抗原设计和莱克多巴胺高敏感性和特异性单克隆抗体的筛选

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摘要

Ractopamine (RCT) is a beta-adrenergic agonist and its use is forbidden in many countries. With the aim of more sensitive and specific detection of RCT, a RCT structure fragment-based hapten was synthesized by introducing a carboxyl group to octopamine through a reductive amination reaction with ethyl levulinate. Six mice were immunized with a hapten–KLH conjugate, and the best monoclonal antibody (Mab) with an IC50 value of 0.34 ng mL?1 in PBST buffer and 0.41 ng mL?1 in a 10-times diluted swine urine sample was obtained, which had no cross reactivity with clenbuterol and salbutamol. These results demonstrated that a fragment of RCT as a hapten can mimic the antigenicity and specificity of RCT very well, and the Mab has potential applications in the specific monitoring of the illegal use of RCT in real samples...
机译:莱克多巴胺(RCT)是一种β-肾上腺素能激动剂,在许多国家/地区都禁止使用它。为了更灵敏,更特异性地检测RCT,通过与乙酰丙酸乙酯的还原胺化反应将羧基引入章鱼胺,从而合成了基于RCT结构片段的半抗原。用半抗原-KLH偶联物免疫六只小鼠,获得最佳单克隆抗体(Mab),在PBST缓冲液中IC50值为0.34 ng mL?1,在10倍稀释猪尿液样品中获得0.41 ng mL?1,与克仑特罗和沙丁胺醇没有交叉反应性。这些结果表明,作为半抗原的RCT片段可以很好地模仿RCT的抗原性和特异性,并且Mab在潜在监控实际样品中非法使用RCT方面具有潜在的应用前景。

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