Em18；多克隆抗血清；制备_

泡球蚴Em18多克隆抗血清的制备和鉴定

http://www.100md.com 《新疆医科大学学报》 2007年第1期

Em18；多克隆抗血清；制备,,Em18；多克隆抗血清；制备,关键词：Em18；多克隆抗血清；制备,1材料和方法,2结果,3讨论,参考文献:

     摘要: 目的：研究泡球蚴18(Em18)抗原的多克隆抗血清的制备和鉴定奠定基础。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，用ELISA和Western印迹分析方法鉴定其特异性。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50 kDa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∶25 600；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。

    关键词： Em18；多克隆抗血清；制备

    Preparation and Indentification of Polyclonal Antiserum against Em18 Antigen

    WANG Junfang, LIN Renyong, ZHANG Chuntao, et al

    (Xingjiang Key Lab of Hydatid Fundamental Medical Research, First Affiliated Hospital, Xinjiang Medical University, Urumqi 830054， China)

    Abstract：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) Obstain high purity rEm18GST recombinant protein 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 ......

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