MA104细胞牛轮状病毒受体的提纯及鉴定

目的研究ＭＡ１０４细胞上的牛轮状病毒（ＢＲＶ）受体。方法ＢＲＶ受体的单克隆抗体（单抗）与溴化氰活化的Ｓｅｐｈａｒｏｓｅ４Ｂ偶联，利用免疫亲和层析法从ＭＡ１０４细胞膜制备物中提纯了ＢＲＶ受体。结果间接酶联免疫吸附试验（ＥＬＩＳＡ）及免疫斑点法表明提纯物呈ＢＲＶ受体抗原阳性；ＳＤＳ－聚丙烯酰胺凝胶电泳显示单抗偶联柱层析洗脱物仅有单一区带与洗脱峰对应；Ｗｅｓｔｅｒｎｂｌｏｔ表明提纯物能够免被受体单抗识别；固相分析实验表明纯化物具有结合ＢＲＶ的能力；提纯物的小鼠免疫血清可以阻断ＢＲＶ感染宿主细胞。结论纯化物为ＢＲＶ受体

Purification and identification of the bovine rotavirus receptor on MA104 cells

In this research the bovine rotavirus(BRV)receptor on MA104 cells was purified and identified.The monoclonal antibody(McAb)against BRV receptor was coupled with the CNBr-activited sepharose 4B. BRV receptor was purified from the MA104 cell extracts by the method of the immunoaffinity chromatography. The results of the indirect ELISA ard the dot immunobindiag assay showed that the BRV receptor antigenicity of the purified compound was positive.On SDS-polyacrylamide gel electrophoresis (SDS-PAGE)the eluate from the column with the McAb coupled showed a single band,corresponding to the elution peak.The eluate from the column without antibody showed no such a band on SDS-PAGE. The result of Western blot showed the purified compound could be distinguished by the McAb against BRVreceptor.Solid-phase assay showed the purified compound had the ability to bind BRV.The mouse immunoserum against the purified compound could block the BRV infection to its host cells. The experimental results above showed that the purified compound was the receptor for BRV.