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重组鼠疫菌F1抗原的纯化及其在ELISA检测鼠疫免疫中的应用

Purification of rF1 antigen expressed in E.Coli and it’s application in detection of anti-plaque response by ELISA

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【作者】 王革; 常娅莉; 万艳; 马超; 王秉翔;

【Author】 WANG Ge,CHANG Ya-li,WAN Yan,et al.(Lanzhou Institute of Biological Products,Lanzhou 730046,China)

【机构】 兰州生物制品研究所; 兰州生物制品研究所 兰州730046; 兰州730046;

【摘要】 从表达rF1抗原的大肠杆菌中以Superdex-200凝胶过滤层析纯化rF1抗原,电泳扫描显示纯化率>90%。SDS-PAGE及琼脂免疫双扩散结果显示纯化的rF1抗原具天然F1抗原的活性。将此纯化的rF1抗原用于间接ELISA分析免疫动物血清中抗F1抗体的水平,并与天然F1抗原相比较,证明rF1抗原优于天然F1抗原分析结果,可用于鼠疫的血清学检测。

【Abstract】 E.Coli DE3-F1STH strain was cultivated with LB.The crude expressed recombinant F1 antigen was further purified by Supherdex-200 Hilode preparative gel filtration chromatography.The purity was reached to more than 90%.After antigen analysis,SDS-PAGE revealed the monomeric form of rF1 with a molecular weight of 17.6kD.Immunodiffusion indicated that the rF1 antigen reacted with the F1-specific antibody.In comparison of the activities with an indirect ELISA in between rF1 and native F1,and the former is superior to the latter.The results demonstrated that rF1 antigen can be used as a diagnostic reagent to quantify the concentration of anti-plague antibody in serum.

【关键词】 重组F1抗原; 纯化; 血清学检测;
【Key words】 rF1 antigen; Purification; Immune serum assay;
  • 【文献出处】 微生物学免疫学进展 ,Progress in Microbiology and Immunology , 编辑部邮箱 ,2006年01期
  • 【分类号】R392
  • 【被引频次】2
  • 【下载频次】116
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