罗汉果甜苷V人工抗原免疫原性检测法的建立与评价

Detection and evaluation of the immunogenicity of Mogrosides V artificial antigen

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。

The study is aimed to prepare antiserum of the Mogroside V-carrier protein complex and determine its immunogenicity,which will be used in the establishment of an enzyme-linked immunosorbent asay（ELISA）-based detection method for MogV and will provide a technical basis for further production of monoclonal antibody against MogV.Firstly,MogV with succinic anhydride,the carrier protein bovine serum albumin（BSA）,and human serum albumin（HSA） were coupled reacted to obtain hapten carrier protein complex;secondly the number of hapten was calculated by UV scanning spectrum method.Then this antigen was used to immunize Balb/c mice to generate antiserum.Finally we used ELISA to detect the titer and specificity of the antiserum.The results showed that the binding ratio of artificial antigen MogV-HS-BSA and MogV-HS-HSA was 44∶1 and 64∶1 respectively;specific antiserum against MogV was obtained by immunization of mice,and the antiserum had high titer.The results indicated that the artificial antigens against MogV have good immunogenicity,and the methods used in this study are successful,simple and feasible.