菜克多巴胺人工抗原的光谱表征及免疫鉴定研究

盐酸菜克多巴胺(Ractopamine hydrochloride,RCT)是一种β-肾上腺素受体激动剂,即使极低剂量残留对人类健康也有危害.为了建立简单、快速并适应现场检测的免疫分析方法,必须先合成它的人工抗原并进行鉴定.通过对莱克多巴胺进行结构修饰,用碳二亚胺将结构修饰了的莱克多巴胺与牛血清蛋白相偶联,经透析得到纯的莱克多巴胺人工抗原并冷冻干燥备用.结果表明,紫外光谱图中合成的人工抗原的特征吸收峰介于蛋白质和莱克多巴胺的特征吸收峰之间;人工抗原红外光谱图中呈现莱克多巴胺和牛血清蛋白的红外特征吸收峰;通过竞争ELISA实验表明用合成的人工抗原免疫的Balb/c小鼠血清中产生了莱克多巴胺的抗体,三种方法综合鉴定结果表明,莱克多巴胺人工抗原合成成功,该人工抗原可以进一步用于制备抗莱克多巴胺的单克隆抗体.

Study on spectral and immune identification of artificial antigen of ractopamine

Ractopamine (RCT) is a β-adrenergic receptor agonist.Because trace residual amount of RCT might be harmful to human being,instrument analysis methods have been developed for the determination of RCT.But these instrument analysis methods require complicated pre-treatment steps and did not suit rapid processing of multiple samples.Immunochromatography assay method can provide in situ monitoring of trace RCT in Foods.In order to build the simple and fast immunochromatography assay method,it is necessary to synthesize and identify artificial antigen of RCT firstly.The modified RCT was conjugated with the carrier of bovine serum albumin (BSA) by EDC· HCI,after dialyzed,the synthesized artificial antigen of RCT was lyophilized, and then stored at-20℃.The results showed that UV spectrum of artificial antigen of RCT had one absorbance band at 276nm which was between the characteristic absorhance band of BSA and RCT.The IR spectrum absorbance bands of BSA appeared on the IR spectrum graph of artificial antigen of RCT, moreover, shrinked vibration bands at 1000-1300cm~1 showed that a C-N bond was produced and suggested that the modified RCT and BSA had been linked.The antibody of anti-RCT was raised in sera of immunized Balb/c mice by indirect competitive ELISA.So the above results showed that the artificial antigen of RCT was synthesized successfully.The artificial antigen of RCT can be used to develop monocIonal antibody of anti-RCT and also to prepare immunochromatography assay method for in situ monitoring of RCT in the future.