抗泡球蚴重组Em18抗原多克隆抗体的纯化与鉴定

Purification and identification of polyclonal antibodies against the recombinant Em18 antigen of Echinococcus multilocularis

目的:制备抗泡球蚴18(Em18)抗原多克隆抗体,纯化并鉴定。方法:表达并纯化rEm18-GST蛋白。用纯化后的蛋白作为免疫原,免疫新西兰白兔获得抗rEm18-GST的多克隆抗体IgG,采用ELISA法检测抗体效价,进一步纯化抗体,获得抗Em18特异性多克隆抗体IgG,用Western blot及ELISA法检测并进行初步鉴定。结果:免疫制备获得兔抗rEm18-GST抗体,随着免疫次数的增加,抗体滴度不断升高,于第10周达到最高。ELISA检测抗体效价为1∶51200。Western blot及ELISA法鉴定表明Em18-GST抗体去除了与GST的交叉反应。结论:获得了去除与GST交叉反应的抗Em18多克隆抗体,为进行噬菌体肽库筛选奠定基础。

Objective： To prepareto purify and identify polyclonal antibodies against recombinant Em18 antigen. Methods： The rEm18-GST fusion protein was expressed and purified using His Bind Resin and the polyclonal antibodies against rEm18-GST were produced by immunizing New-Zealand white rabbits. The titer of polyclonal antibody was detected by ELISA and then the purified specific anti-Em18 polyclonal antibodies were gotten and were checked by Western blot and ELISA. Results. The titer increased during weeks and was 1 ： 51 200 in the end. The crossing reaction with GST was deleted according to the Western blot and ELISA. Conclusion. The anti Em18 polyclonal antibodies which do not have the crossing reaction with GST were gotten.