三聚氰胺人工抗原的合成与鉴定

Synthesis and Identification of Artificial Antigen for Melamine

目的:建立三聚氰胺残留的免疫分析方法。方法:以三聚氰胺结构类似物2-氯-4,6-二氨基-1,3,5-三嗪为原料,分别与6-氨基己酸和对氨基苯甲酸,各经两步化学反应合成两种具有不同长度间隔臂的半抗原MEA和MEB,并分别通过碳二亚胺法和混合酸酐法将MEA和MEB与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,制备三聚氰胺的免疫原(MEA-BSA)和包被原(MEB-OVA)。结果:经质谱和紫外光谱表征,证明所合成的产物为目标半抗原MEA和MEB,并且与载体蛋白偶联比可达到25.28:1(MEA-BSA)和13.11:1(MEA-OVA)。结论:半抗原MEA和MEB及人工抗原合成成功。

To develop an immunoassay for melamine residues,melamine hapten MEA and MEB were synthesized by the reaction between 2-Chloro-4,6-diamino-1,3,5-triazine and aminocaproic acid or p-aminobenzoic acid,respectively.Then two artificial antigens（MEA-BSA and MEB-OVA） were prepared by active ester method（AE） and mixed anhydride method（MA） respectively.Mass spectral analysis demonstrated the successful synthesis of MEA and MEB.Characterization of MEA-BSA and MEB-OVA was conducted by using UV spectrophotometric method.The numbers of hapten molecules per protein molecule estimated by ultraviolet spectrophotometry according to the absorbance at 253.5 nm for two conjugates were 25.28 and 13.11 respectively.The results suggested that the artificial antigens can be used for preparing specificity anti-melamine antibody.