甘草酸人工抗原的合成鉴定及免疫原性分析

目的制备中药甘草的活性成分甘草酸(GA)的人工抗原及抗血清,为制备GA的单克隆抗体、并建立快速检测GA的酶联免疫吸附测定(ELISA)提供技术基础。方法将GA与载体蛋白牛血清白蛋白(BSA)偶联起来制得完全抗原后,经基质辅助激光解吸飞行时间质谱鉴定其相对分子质量,用此抗原免疫BALB/c小鼠,制备抗血清,并通过间接ELISA法和竞争ELISA法检测其抗体效价和特异性。结果合成的人工抗原GA-BSA中GA与BSA的结合比约为7∶1;免疫小鼠得到特异针对GA的多抗血清,GA抗体的效价为1∶8000。结论成功地合成了GA的人工抗原,且该抗原有较好的免疫原性,可应用于建立GA的免疫分析方法。

Identification and immunogenicity analysis of artificial antigen of Glycyrrhizic acid conjugated with BSA

Purpose To prepare the conjugate antigen Glycyrrhizic acid-bovine serum albumin(GA-BSA) and its antiserum to produce anti-GA monoclonal antibody and to establish a fast assay for GA content by an ELISA.Methods The conjugate antigen GA-BSA was artificially coupled and identified by matrix-assisted laser absorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS).BALB/c mice were immunized with GA-BSA to prepare antiserum against GA.The antiserum titer and specificity were detected by indirect ELISA and competitive ELISA,respectively.Results The conjugating ratio of GA and carrier BSA was 7∶1.The antiserum titer was 1∶8 000 and it reacted specifically to the GA.Conclusion The successfully synthesized conjugate antigen GA-BSA implies its feasibility in the establishment of fast immunoassay for the GA content determination