【摘要】 目的:制备抗水痘-带状疱疹病毒(VZV)主要衣壳蛋白ORF40的单克隆抗体,对该蛋白在VZV感染细胞内的表达、分布以及在VZV病毒颗粒中的包含情况进行初步探究。方法:设计并合成VZV ORF40多肽片段,偶联牛血清白蛋白后免疫小鼠,制备与筛选抗VZV ORF40单克隆抗体,使用获得的抗体通过Western blot和免疫荧光法对VZV感染细胞进行检测,并通过Western blot对纯化的VZV病毒颗粒进行检测。结果:获得VZV ORF40单克隆抗体10F2,该抗体可特异性识别VZV感染细胞表达的以及VZV病毒颗粒包含的ORF40蛋白;本研究应用该抗体确定了ORF40蛋白在VZV感染细胞中的核定位,并且发现ORF40蛋白在核周有点状聚集现象,在细胞质区域偶尔也能检测到ORF40蛋白的分布。结论:本研究筛选获得抗VZV ORF40的特异性单克隆抗体,为后续开展该蛋白在VZV感染与病毒颗粒组装过程中的功能研究奠定基础。更多还原

【Abstract】 Objective: To prepare monoclonal antibodies against ORF40,the major capsid protein of varicella-zoster virus( VZV),and to investigate the expression and distribution of ORF40 in VZV infected cells and its inclusion in VZV particles. Methods:The polypeptide antigen of VZV ORF40 was designed,synthesized and conjugated to bovine serum albumin. After immunization in mice,the monoclonal antibodies( m Abs) against VZV ORF40 were screened and prepared. Using anti-VZV-ORF40 m Abs,VZV-infected cells were analyzed by Western blot and immunofluorescence,while purified VZV particles were analyzed by Western blot. Results: An anti-VZV-ORF40 m Ab 10 F2 was obtained,which specifically recognized ORF40 that expressed in VZV-infected cells or contained in VZV particles. Using this antibody,the nuclear localization of ORF40 protein in VZV-infected cells was determined,and a distinct punctate perinuclear pattern of ORF40 localization was observed. Moreover,the distribution of ORF40 protein was occasionally detected in the cytoplasm. Conclusion: In this study,a specific anti-VZV-ORF40 m Ab was obtained,and this m Ab could lay a foundation for further study on the function of ORF40 in the process of VZV infection and virus particle assembly.更多还原