氨基脲人工抗原的合成及其多克隆抗体的制备

目的合成氨基脲人工抗原并制备其多克隆抗体。方法以氨基脲(SEM)和对醛基苯甲酸(CP)为原料合成半抗原(CP-SEM),并采用碳化二亚胺法和混合酸酐法将其分别与牛血清白蛋白(BSA)和鸡卵清蛋白(OVA)偶联,合成氨基脲人工抗原。以氨基脲人工抗原(CPSEM-BSA)为免疫原免疫Balb/c小鼠,利用间接ELISA法测定其抗体效价。结果经薄层层析、红外光谱和元素分析等方法鉴定合成半抗原即为CP-SEM。紫外扫描、聚丙烯酰胺凝胶电泳结果表明人工抗原合成成功。免疫获得抗氨基脲的多克隆抗体,其抗体效价为1:64000。结论氨基脲人工抗原合成成功,并获得抗氨基脲的多克隆抗体。

Synthesis of Semicarbazide artificial antigen and Preparation of Its Multi-Clonal Antibody

Objective To produce the artificial antigen and multi-clonal Antibody of Semicarbazide(SEM).Methods Firstly,the hapten(CP-SEM) was derivatived by combining 4-carboxybenzaldehyde(CP) with Semicarbazide(SEM).Secondly,the hapten was conjugated to bovine serum albumin(BSA) and ovalbumin(OVA) by carbodiimide method and the mixed anhydride method respectively.Finally,the artificial antigen(CPSEM-BSA) was used to immunized Balb/c mice to obtain polyclonal antibodies.the antibody to SEM were determined by indirect ELISA.Results:SEM artificial antigens were synthesized successfully,which were proved by ultraviolet spectrum and SDS-PAGE.Test of the antisera by indirected ELISA showed that the titer was 1:64000.Conclusion The results showed that the synthesis of antigen and preparation of multi-clonal antibody against SEM were successful.