抗HBsAg单克隆抗体的制备与生物学特性的鉴定  

Preparation and characterization of monoclonal antibody against S protein of HBV

作　　者：王雪梅[1] 赖梅梅[1] 张晓丽[2] 张祯祯[1] 陈婧[1] 张君[1] 黄爱龙[1] 

机构地区：[1]重庆医科大学附属第二医院病毒性肝炎研究所感染性疾病分子生物学教育部重点实验室,重庆400016 [2]第三军医大学附属西南医院肿瘤科,重庆400038

出　　处：《重庆医科大学学报》2008年第3期267-269,共3页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助(项目号：30400374)。

摘　　要：目的：制备分泌抗HBVS蛋白的单克隆抗体的杂交瘤细胞株，并鉴定其特性，为建立快速诊断HBV感染的方法奠定基础。方法：以重组乙肝疫苗免疫BALB／c小鼠，通过细胞融合建立了能稳定分泌抗HBsAg单抗的杂交瘤细胞株，利用间接ELISA技术和Western blot进行单抗特异性的鉴定，同时采用间接ELISA方法鉴定mAb的Ig亚类，检测mAb的效价及相对亲和力。结果：获得1株可分泌特异性mAb的杂交瘤细胞4D2，其抗体亚类为IgG1型，腹水效价为1：106，相对亲和力在10^5以上。结论：成功地制备出抗HBVS蛋白的单克隆抗体，为建立快速特异检测HBV感染的实验方法提供了有力的工具。Objective：To prepare anti-HBV S protein hybridoma cell line, this will be helpful to the establishment of rapid HBV infection diagnosis assay. Methods：BALB/c mice were immunized with the recombinant Hepatitis B vaccine. A hybridoma cell line which consistently secreted monoclonal antibody（McAb）against HBsAg was obtained through cell fusion. The specificity of the McAb was analysed by indirect ELISA and Western blot analysis.The immunoglobulin （Ig） subtypethe ascites titers, and the affinity of the obtained mAbs were determined by indirect ELISA. Results：From over hundred positive hybridomas which secreted anti-S protein mAbs,one of hybridomas was screened out,designated 4D2. The subtype of the McAb was IgG1. The titer of 4D2 mAb produced by ascites fluid was over 1 ： 106. The relative affinity constant of 4D2 mAb was determined as 10^5. Conclusion：One of high titer,specific mAb against HBV S protein has been successfully prepared and primarily identified,which may be useful in the development of a rapid and convenient diagnostic kit for detection of HBV infection.

关 键 词：乙型肝炎病毒 乙型肝炎表面抗原 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学]