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Specific Depletion of Naringin from Si-Ni-San, a Traditional Chinese Prescription, by an Immunoaffinity Chromatography
Author(s): 
Pages: 243-246
Year: Issue:  3
Journal: CHINESE JOURNAL OF CHROMATOGRAPHY

Keyword:  免疫亲和色谱柚皮苷完全抗原特异性剔除四逆散;
Abstract: 为了获得剔除柚皮苷(naringin)的中药方剂四逆散样品以供其药理活性探讨时使用,制备了抗柚皮苷抗体的免疫亲和色谱柱,用于特异性地剔除四逆散中的柚皮苷.首先合成了柚皮苷的完全抗原柚皮苷与牛血清白蛋白的结合物naringin-BSA,并用naringin-BSA对新西兰兔进行免疫获得抗血清,再将其纯化后与经CNBr活化的Sepharose 4B凝胶共价偶联制成免疫亲和色谱柱.将四逆散提取物样品溶液上样该色谱柱,洗脱,制得特异性剔除了柚皮苷的四逆散样品.由检测结果可知,naringin-BSA被成功合成.将其用于免疫新西兰兔,获得的抗血清的效价经酶联免疫吸附法(ELISA)测定达到1∶30000,抗体IgG的纯度达94% ,交叉反应率低.在IgG与Sepharose 4B合成的IgG-Sepharose免疫亲和色谱柱中,IgG的偶联率为87% .用该免疫亲和色谱柱处理四逆散后,其中所含的柚皮苷几乎完全被剔除.结果证明,利用抗柚皮苷免疫亲和色谱,能特异性地剔除四逆散或其他样品中的柚皮苷成分.
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