目的:制备抗猪瘟病毒(CSFV)C株 E~(rns)/E2嵌合基因表达蛋白的单克隆抗体(mAb),探讨~(rns)/E2蛋白的分子结构和生物学功能.方法:在大肠杆菌 Rosetta~(TM)2(DE_3)中表达了猪瘟病毒(CSFV)C株 E~(rns)、E2蛋白主要抗原域的嵌合基因CSFV E~(rns)/E2蛋白;以纯化复性后的CSFV E~(rns)/E2为免疫原,按常规方法免疫BALB/c小鼠,应用淋巴细胞杂交瘤技术制备抗CSFV E~(rns)/E2 mAb;利用间接ELISA 、Western blot、Dot blot方法鉴定mAb生物学特性.结果:获得了1株稳定分泌抗CSFV Erns/E2融合蛋白的单克隆杂交瘤细胞系C8株,该株mAb的细胞培养上清效价为 1:6 400,小鼠腹水效价 1:2×10~5.其免疫球蛋白亚类为IgG1.间接ELISA和Western blot结果显示该株mAb不与猪瘟C株细胞毒、pET-32a标签蛋白反应,仅与融合表达的 CSFVE~(rns)/E2蛋白存在特异性反应.Dot blot结果表明其识别的抗原表位为E2蛋白的 RSGLCPFDTSPV氨基酸序列.结论:获得了一株能特异识别E2蛋白抗原表位的 mAb,为进一步研究 CSFV 的分子结构及功能奠定了物质基础.

To prepare hyperimmunized rabbit serum against classical swine fever virus(CSFV) C strain,the Zelanian rabbit were immunized with the attenuated CSFV vaccine.The serum titer of immunized rabbit was determined by indirect ELISA and the IgG was purified by saturated ammonium sulfate precipitation.Then,the purity of IgG was checked by SDS-PAGE electrophoresis and the safety was confirmed by microorganism test and inoculating piglets.The results showed that the protein concentration of the purified IgG was 6 mg/mL,and the ELISA titer was 1∶6400.Taken together,we have produced serum IgG against CSFV,which shows high titer,high safety,low cost,and can be used to treat or prevent swine fever.It is an ideal biological agent for the detection of CSFV and the control of CSF.

综述了国内外IBDV细胞受体的研究情况,概括介绍了近年来的研究进展,以期对IBDV细胞受体的结构和功能研究提供参考.