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目的制备乙酰胆碱脂酶抑制活性成分石杉碱甲(huperzine A,HupA)的人工抗原及抗血清。方法将HupA与戊二醛、载体蛋白牛血清白蛋白(BSA)反应偶联,制得半抗原-载体蛋白复合物后,用基质辅助激光解吸飞行时间质谱测定其中结合的半抗原数目;以此抗原免疫BALB/c小鼠,制备抗血清,并通过ELISA法检测其抗体效价和特异性。结果合成的人工抗原HupA-GA-BSA中HupA与BSA的结合比约为8∶1;免疫小鼠得到特异针对HupA的抗血清,HupA抗体的效价为1∶62500。结论成功地合成了HupA的人工抗原,且该抗原有较好的免疫原性。
Objective To prepare artificial antigen and antiserum of huperzine A (HupA), an active ingredient of acetylcholinesterase. Methods The hapten-carrier protein complex was prepared by coupling HupA with glutaraldehyde and carrier protein bovine serum albumin (BSA), and then the number of hapten bound was determined by matrix-assisted laser desorption / emission time-of-flight mass spectrometry BALB / c mice were immunized with antigen to prepare antiserum, and their antibody titer and specificity were tested by ELISA. Results The binding ratio of HupA to BSA in synthetic human antigen HupA-GA-BSA was about 8: 1. The antiserum specific to HupA was obtained from the immunized mice. The titer of HupA antibody was 1: 62 500. Conclusion HupA artificial antigen was successfully synthesized, and the antigen has good immunogenicity.