泡球蚴Em18多克隆抗血清的制备和鉴定

Preparation and indentification of polyclonal antiserum against Em18 antigen

目的:制备和鉴定泡球蚴Em18抗原的多克隆抗血清。方法:利用大肠杆菌BL21(DE3)表达重组质粒pET41a-Em18,经异丙基硫代-β-D-半乳糖苷(IPTG)诱导、表达和纯化rEm18-GST蛋白,免疫BALB/c小鼠,制备鼠源抗Em18多克隆抗体,采用ELISA和Western blot法鉴定其特异性。结果:(1)SDS-PAGE检测表明,rEm18-GST蛋白得到成功表达,在相对分子量为50kDa处有表达条带。(2)纯化后获得高纯度的rEm18-GST蛋白。(3)ELISA结果表明,抗Em18抗血清的抗体滴度为1∶25600;Western blot结果表明抗Em18抗血清能与rEm18-GST蛋白发生特异性结合。结论:获得了特异性识别rEm18-GST蛋白的多克隆抗体,为进一步筛选Em18模拟抗原表位奠定基础。

Objective. To obstain polyclonal antiserum against Em18 antigen for the foundation of further screening for Em18 related peptides. Methods： The recombinant expression plasmid pET41a-Em18 was transformed into BL21 （E. coli） via IPTG induction and purified rEm18-GST as immunogen was used to immunize BALB/c mice. The specificity of mice anti-Em18-GST antibody was analyzed by Western blot and ELISA. Results： （1） The expressed rEm18-GST recombinant protein can be detected as a band of 50 kDa by SDS-PAGE. （2） High purity rEml8-GST recombinant protein was obtained after purification. （3） The polyclonal anti-Em18-GST antibody could specifically recognize rEm18-GST recombinant protein. ELISA showed a high titre （1 ： 25 600） of the antiserum. Conclusion. Polycloned anti-Em18-GST antibody with high specificity and purity has been prepared by using purified rEm18-GST as immunogen, which lays the foundation for further screening for Em18 related peptides.