摘要
目的：合成芒果苷人工抗原并进行免疫原性鉴定。方法：采用经典高碘酸钠法,将芒果苷（mangiferin,MRn）分别与牛血清白蛋白（BSA）和鸡卵清白蛋白（OVA）相耦联合成MRn的人工抗原和包被原。用紫外扫描的方法测定其耦联比,并免疫动物,通过间接ELISA方法测定芒果苷抗体的效价,采用间接竞争ELISA（ic ELISA）法鉴定芒果苷人工抗原（MRn-BSA）的免疫原性。结果：MRn-BSA的耦联比为6∶1,ic ELISA实验结果表明,4只人工抗原MRn-BSA免疫的小鼠均有特异性抗体产生,抗体效价：1∶4 000以上,线性检测范围为1~100μg/m L。结论：该方法成功合成了MRn的人工抗原,并且在免疫小鼠的血清中产生了针对MRn的抗体,为下一步芒果苷的单克隆抗体制备以及相应免疫方法的建立奠定基础。
Objective： To establish an enzyme linked immunosorbent assay（ELISA） for mangiferin（MRn）,artificial antigen of MRn was synthesized. Methods： Oxidation method using sodium iodide was used to synthesize immunogenic antigen（MRn-BSA） and coating antigen（MRn-OVA） of MRn. The characterization of the synthesis was examined by UV spectrometry. BALB/c mice were immunized with the prepared MRn-BSA immunogenic antigen. The titer of the anti-serum was detected by ELISA,in the meanwhile the immunogenicity of MRn-BSA was confirmed by indirect competitive ELISA（ic ELISA）. Results： UV spectroscopy showed that MRn was successfully conjugated with OVA and BSA,so the new compound of MRn-BSA was synthesized,the coupling ratio was 6∶ 1. After immuned MRn-BSA,the mice could produce anti-MRn antibodies specifically,of which titer was up to 1∶ 4 000,and the general range was1 ~ 100 μg/m L. Conclusion： Anti-MRn antibodies are discovered in the serum of mice,which indicates that artificial antigen of MRn is successfully synthesized,for the purpose of preparing monoclonal antibodies,as well as the establishment of appropriate immune methods.
出处
《中药材》
CAS
CSCD
北大核心
2015年第1期119-122,共4页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(81373542
81274043)
关键词
芒果苷
人工抗原
高碘酸钠法
免疫分析法
抗芒果苷抗体
Mangiferin
Artificial antigen
Sodium periodate oxidation method
Immunoassay methods
Anti-mangiferin antibodies
