摘要
目的合成石杉碱甲人工抗原并进行鉴定。方法采用戊二醛法,将石杉碱甲与牛血清白蛋白偶联,通过紫外光谱扫描和SDS-PAGE凝胶电泳鉴定偶联是否成功,根据摩尔分子比公式测定其偶联比,并通过L9(33)正交试验考察反应物起始摩尔比、反应时间和戊二醛加入量对偶联率的影响。结果人工抗原的紫外光谱扫描与半抗原和载体蛋白的紫外光谱相比均发生变化;人工抗原SDS-PAGE凝胶电泳条带与载体蛋白相比发生偏移;半抗原与载体蛋白的偶联比为10.8∶1。结论该方法成功合成石杉碱甲的人工抗原,并且确定戊二醛法的最佳反应条件为:反应物起始摩尔比100∶1,反应时间8 h,戊二醛加入量80μL。
Objective To synthetize and identify the artificial antigens of huperzine A. Methods Huperzine A was coupled with Bovine Serum Albumin （BSA） by the method of glutaraldehyde, and the couple was identified by UV scan and SDS-PAGE electrophoresis. The coupling ratio was detected by the molar ratio formula, and some influence factors such as the initial molar ratio of reagents, the coupling time and the addition of coupling agent to the coupling ratio were investigated by L9 （33） orthogonal design test. Results The results of UV scan of artificial antigens were changed compared with those of hapten huperzine A and carrier protein. SDS-PAGE electrophoresis strip of artificial antigens changed compared with that of carrier protein. The coupling ratio of huperzine A to BSA was 10.8：1. Conclusion The artificial antigens of huperzine A can be synthetized successfully by glutaraldehyde, and the best factors are as follows： the initial molar ratio of reagents is 100：1, the coupling time is 8 hours and the addition of glutaraldehyde is 80μl.
出处
《福建中医药大学学报》
2012年第6期41-44,共4页
Journal of Fujian College of Traditional Chinese Medicine
基金
福建省科技厅重点项目(2011Y0035)
福建省科技计划项目(2010Y2004)
关键词
石杉碱甲
人工抗原
戊二醛
偶联率
huperzine A
artificial antigens
glutaraldehyde
coupling ratio
作者简介
滕海英（1988-），女，2010级药物分菥专业硕士研究生，主要从事药物分析研究。
通讯作者：余宇燕（1973-），女，理学博士，副教授，硕士师导师。E-mail：yyyfj@163．com
