抗雌激素多克隆抗体的制备及效价鉴定

Preparation of Anti-estrogen Polyclonal Antibody and Identification of Its Titer

为了快速安全地检测动物性食品中是否含有过高浓度雌激素,本研究将半抗原雌二醇(Estradiol,E_(2))合成为雌二醇-3-羧甲基醚(E_(2)-3-CME),再通过碳二亚胺(EDC)法与牛血清白蛋白(BSA)结合,合成完全抗原(E_(2)-BSA).将1 mg/只的E_(2)-BSA免疫雌性家兔,制备出抗雌激素的抗体.通过用酶联免疫吸附(Elisa)实验对产生的抗体进行效价鉴定.实验结果表明,第三次免疫后E_(2)-BSA产生的抗体光密度值(OD)最高,且与对照组,免疫一次和两次产生的抗体的OD值均具有明显差异,显示通过抗原免疫三次的家兔产生的抗体效价最高,可为后期通过此方法制备抗体和Elisa试剂盒奠定基础.

In order to quickly and safely detect whether there are excessive concentrations of estrogen in animal foods,hapten estradiol(Estradiol,E)was synthesized into estradiol-3-carboxymethyl ether(E-3-CME),and then combined with bovine serum albumin(BSA)to be synthesized complete antigen(E-BSA)by the carbodiimide(EDC)method.Female rabbits were immunized with E-BSA of 1 mg/rabbit to prepare anti-estrogen antibodies.The titer of antibody was identified by using enzyme-linked immunosorbent assay(Elisa).The experimental results showed that the optical density(OD)value of the antibody produced by E-BSA was the highest after the third immunization,and it was significantly different from these groups of control,once and twice after immunization.The conclusion shows that the antibody titer produced by rabbits immunized with antigen for three times is the highest,which can lay a foundation for the preparation of antibody and Elisa kit by using this method in the later stage.