论文部分内容阅读
目的制备罗汉果甜苷V(mogroside V,MogV)-载体蛋白复合物人工抗原及抗血清,建立抗血清与MogV特异性反应的检测方法,为进一步制备MogV的单克隆抗体、建立快速检测MogV的ELISA法提供技术基础。方法将MogV与丁二酸酐、载体蛋白牛血清蛋白(BSA)和人血清蛋白(HSA)反应偶联,制得半抗原-载体蛋白复合物后,通过紫外扫描光谱计算出结合的半抗原数目;以此抗原免疫Balb/c小鼠,制备抗血清,并通过ELISA法检测效价和特异性。结果合成的人工抗原MogV-HS-BSA结合比约为44∶1,MogV-HS-HSA结合比约为64∶1;通过免疫小鼠得到特异性针对MogV的抗血清,血清效价较高。结论 MogV人工抗原有较好的免疫原性,建立的免疫原性检测方法简便可行。
Objective To prepare artificial antigen and antiserum of mogroside V (MogV) - carrier protein complex and to establish a method for detecting the specific reaction between antiserum and MogV. To further prepare a monoclonal antibody against MogV, an ELISA Law provides the technical basis. Methods MogV was conjugated with succinic anhydride, BSA and HSA to obtain hapten - carrier protein complex, and then the number of hapten bound was calculated by UV - scanning spectroscopy. Balb / c mice were immunized with this antigen, antisera were prepared, and titer and specificity were determined by ELISA. Results The binding ratio of MogV-HS-BSA was about 44:1. The binding ratio of MogV-HS-HSA was about 64:1. The antiserum specific to MogV was obtained by immunizing mice with high serum titer. Conclusion MogV artificial antigen has good immunogenicity, and the established immunogenicity test method is simple and feasible.