高价特异半抗原雌二醇抗体及酶标雌二醇复合物的研制

Preparation and Application of Antiestradiol Antobodies and Enzyme Labeled Estradiol

目的：制备出高价特异的半抗原雌二醇抗体和高纯度稳定的雌二醇－１７半琥珀酸－辣根过氧化物酶（ＨＲＰ）复合物。方法：主要使用材料包括半抗原载体复合物、半抗原活化体、ＨＲＰ及免疫用新西兰纯系家兔。具体方法为：（１）自行设计小分子半抗原的微量快速免疫方案，免疫全过程半抗原雌二醇的总使用体重量为３６７．５μｇ／ｋｇ，免疫时间为１１周。（２）首次建立活化４Ｂ珠与ＢＳＡ交联的反相快速亲合免疫吸附技术，以除去抗载体抗体，批次特异纯化了抗雌二醇抗体，（３）改进皮质醇－１７半琥珀酰胺－ＨＲＰ的标记法，建立了雌二醇－１７半琥珀酸－ＨＲＰ标记技术。结果：研制出高价特异的抗雌二醇抗体，效价１∶５万以上、抗体的活性达５ｍｇ／Ｌ；抗载体ＢＳＡ抗体的含量为０。首次制备出高纯度稳定的雌二醇－１７半琥珀酸－ＨＲＰ复合物；用ＥＬＩＳＡ法与自制固相Ｅ２抗体结合鉴定，复合物经ＳｅｐｈａｄｅｘＧ７５纯化后Ａ４５０峰值与Ａ４０３及Ａ２８０基本吻合，复合物的活性为５ｍｇ／Ｌ。结论：本研究建立的半抗原雌二醇抗体及酶标雌二醇复合物的制备、纯化及分析鉴定技术简便、快速经济。

オbjective: To prepare specific antiestradiol antibodies and enzyme labeled estradiol (E2HRP). Methods: Fast and trace immunity. Estradiol: 3675 μg/kg. Fast affinity absorption to move BSA antibodies. Enzyme labeled E2～17hemisuccinateHRP. Results: The titer of antiestradiol antibodies reached 1∶50000～120000 as measured by ELISA. AntiBSA content was zero. The activities of antibodies were 5 mg/L. The activities of E2HRP were 5 mg/L. There were no cross reactions between antiestradiol antibodies and other 4 kinds of steroid substances.