HEV部分ORF_2重组蛋白的纯化及其在酶标试剂中的初步应用  

Purification of Recombinant HEV Partial ORF_2 Protein and Application of It in Preparation of ELISA Kit

作　　者：王玉梅[1] 刘彦彬[1] 李秀芝[1] 刘欣荣[1] 蒙冲[1] 刘畅[1] 郭凤云 

机构地区：[1]长春生物制品研究所,长春130062

出　　处：《中国生物制品学杂志》2001年第3期174-175,共2页Chinese Journal of Biologicals

摘　　要：目的 获得具有生物学活性的重组蛋白，用于制备抗－ＨＥＶ酶标试剂盒。方法 将插入的含ＨＥＶ 部分开放读码框２（ＯＲＦ２）的重组菌扩大培养，诱导表达，纯化，鉴定，组装酶标试剂盒。结果 纯化的重组蛋白经 ＳＤＳ－ＰＡＧＥ鉴定，在相对分子质量５６０００处有一明显的蛋白带，经ＣＳ－９３０薄层扫描纯度达９５％以上，经免疫印迹 分析，具有良好的免疫学特异性。此ＨＥＶ试剂盒，检测阳性病人血清样品阳性率为９４．７４％，检献血员血清样品阳 性率为５．４３％。结论 该纯化的ＨＥＶ部分ＯＲＦ２重组蛋白组装的试剂盒特异性及敏感性均较好。Objective To obtain the recombinant HEV partial ORE2 protein with biological activity and use it for the preparation of anti-HEY ELISA kit. Methods The recombinant F. coli NM522 containing HEY partial ORE2 was cultured, and the expression of ORE2 protein was induced. The expressed product was purified, identified, and used for preparing ELISA kit. Results A single band with a relative molecular weight of 56000 was observed on the SDS-PAGE profile of expressed product after being purified. CS-930 thin layer scanning proved that the purity of it reached above 95% . Western blot showed good immunological specificity of the expressed product. The kit prepared with the expressed protein was used for detecting the serum specimenerns from the patients and blood donors. The result showed that the HEY positive rates of them were 94.74% and 5.43% respectively. Conclusion The kit prepared with recombinant HEY partial ORE2 protein showed specificity and sensitivity.

关 键 词：戊型肝炎病毒 重组蛋白 酶免疫吸附试验 应用 纯化 

分 类 号：R392－33[医药卫生—免疫学]