Synthesis of huperzine A-bovine serum albumin conjugate and its immunogenicity in mice

宋建武; 晁志; 董为人

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Synthesis of huperzine A-bovine serum albumin conjugate and its immunogenicity in mice

更新时间：2026-01-09

- Synthesis of huperzine A-bovine serum albumin conjugate and its immunogenicity in mice
- Journal of Army Medical University Vol. 31, Issue 21, Pages: 2128-2130(2009)
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  CLC： R284.3;R392-33;R392.7
- Published：2009
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宋建武,晁志,董为人.石杉碱甲人工抗原的合成、鉴定及免疫原性分析[J].陆军军医大学学报(原第三军医大学学报),2009,31(21):2128-2130.

　SONG Jian-wu,CHAO Zhi,DONG Wei-ren.Synthesis of huperzine A-bovine serum albumin conjugate and its immunogenicity in mice[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(21):2128-2130.
宋建武,晁志,董为人.石杉碱甲人工抗原的合成、鉴定及免疫原性分析[J].陆军军医大学学报(原第三军医大学学报),2009,31(21):2128-2130. DOI：

　SONG Jian-wu,CHAO Zhi,DONG Wei-ren.Synthesis of huperzine A-bovine serum albumin conjugate and its immunogenicity in mice[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(21):2128-2130. DOI：

摘要

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∶62 500。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。

Abstract

Objective To prepare huperzine A-carrier protein conjugate and determine its immunogenicity for later production of monoclonal antibody against HupA and establishing an enzyme-linked immunosorbent assay (ELISA)-based detection method for HupA. Methods HupA was conjugated with bovine serum albumin (BSA) using glutaraldehyde (GA) method to obtain the HupA-GA-BSA conjugate

and the hapten number in the conjugate was determined by matrix-assisted laser absorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). BALB/c mice were immunized with HupA-GA-BSA to prepare the antiserum against HupA. The serum titer and specificity of the HupA antibodies were detected by indirect ELISA and competitive ELISA

respectively. Results The conjugation ratio of HupA and the carrier protein BSA was 8∶1. The antiserum against HupA with a titer of 1∶62 500 was obtained after immunizing the mice with the conjugate

and the antiserum reacted specifically to HupA. Conclusion The synthesized HupA-GA-BSA conjugate possesses good immunogenicity in mice

suggesting the feasibility of preparing the monoclonal antibody against HupA using this conjugate.

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