2种石房蛤毒素完全抗原交联方法的比较及多克隆抗体的制备

目的]探究石房蛤毒素(STX)完全抗原制备方法和STX多克隆抗体免疫方案。方法]通过碳二亚胺法(EDC)和高碘酸盐法(periodate reaction)2种交联方法,将小分子石房蛤毒素与牛血清白蛋白(BSA)、鸡卵清蛋白(OVA)和孔血蓝蛋白(KLH)分别进行交联,制备了6种形式STX完全抗原,并对交联物进行琼脂糖凝胶电泳鉴定和紫外吸收峰迁移变化鉴定。分别将EDC法和高碘酸盐法交联的STX-BSA、STX-KLH 4种完全抗原作为免疫原,对Balb/c小鼠进行免疫,获得STX多克隆抗体。通过间接ELISA法,对不同方法制备的多克隆抗体进行分析比较。结果]在石房蛤毒素完全抗原的制备中,在交联方法的选择上,EDC法较高碘酸盐法更具优势;而在免疫原的选择上,STX-BSA完全抗原效果最好。结论]本研究探究了2种制备STX完全抗原的方法,为今后多克隆抗体生产以及特异性单克隆抗体筛选提供数据支撑。

Comparison of two complete antigen conjugation methods for saxitoxin and preparation of polyclonal antibody

Aim] In order to explore the preparation method of complete antigen of saxitoxin (STX) and the immunization project of STX polyclonal antibody. Method] We conjugated small molecule STX with bovine serum albumin (BSA) and porocylocyanin (KLH) by carbodiimide method (EDC) and periodate reaction, respectively. Six forms of STX complete antigens were prepared, and the resulted products were identified by agarose gel electrophoresis and UV spectra. Balb/c mice were immunized with STX-BSA and STX-KLH crosslinked by EDC and periodate reaction to induce polyclonal antibodies against STX, and then the titer of the obtained polyclonal antibodies by different methods were analyzed and compared by indirect ELISA. Result] The EDC method is superior to periodate reaction in the selection of conjugation method in the preparation of complete antigen of STX; in the selection of immunogens, STX-BSA complete antigen is the best. Conclusion] This study explored two methods to prepare STX complete antigens, which provided datas support for the production of polyclonal antibodies and the screening of specific monoclonal antibodies in the future.