筛选出一株可识别多种变异HBsAg的单克隆抗体及其应用  

Screening and application of a monoclonal antibody against variant hepatitis B surface antigen

作　　者：田拥军[1] 张振华[1] 李磊[1] 刘慎沛[2] 徐飏[3] 陆蒙吉[3] 杨东亮[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院临床免疫研究室,武汉430030 [2]华中科技大学同济医学院附属同济医院实验医学研究中心,武汉430030 [3]华中科技大学同济医学院病原生物学系,武汉430030

出　　处：《免疫学杂志》2008年第1期56-60,共5页Immunological Journal

基　　金：国家十五科技攻关项目(2001BA705B05);湖北省科技攻关项目(2006AA301B51-1)资助

摘　　要：目的制备筛选可识别变异表面抗原(hepatitis B surface antigen,HBsAg)的单克隆抗体(monoclonal antibody,mAb)。用筛选出的单克隆抗体建立检测变异HBsAg的ELISA实验方法。方法用血源HBsAg免疫Balb/c小鼠,通过杂交瘤细胞融合技术制备抗-HBs单克隆抗体。不同单克隆抗体包被酶标反应孔,检测真核细胞表达的野生及变异HBsAg,了解各种单克隆抗体的反应模式。筛选出可以较好识别变异HBsAg的单克隆抗体Hb1,优化该抗体ELISA检测HBsAg的方法,与8种HB-sAg检测试剂比较检测变异HBsAg的能力。结果经过筛选,得到一种可以较好识别包括G145R在内大多数变异HBsAg的单克隆抗体。检测变异HBsAg的能力优于市售HBsAg诊断试剂。结论用本实验制备的单克隆抗体可以用于ELISA检测变异HBsAg,减少HBsAg变异株的漏检率。Objective To produce monoclonal antibodies （mAbs） that had the ability to detect the variant forms for developing Enzyme-linked immunosorbent assay （ELISA） method of detecting hepatitis B surface antigen mutants using mAb. Methods Babl/e female mice were immunized with HBsAg isolated from serum of HBV infected patient. All monoclonal antibodies were produced according to standard meth- ods. Total of 15 different HBsAg variants were produced in a mammahan expression system. The reactivity of anti-hepatitis B surface antigen monoclonal antibodies to mutant antigens was measured by enzyme immunoassay. The immunoreactivity to mutant antigens was also tested using 8 commercial HBsAg diagnostic assays. Results Total of 14 anti-HBs monoclonal antibodies were produced in the study. One anti-HBsAg monoclonal antibody（Hbl）was selected which could reacted strongly with most tested variants including G145R mutation. Conclusion The Hbl is a valuable tool for detection of HBsAg variants.

关 键 词：乙型肝炎病毒 表面抗原 单克隆抗体 

分 类 号：R392[医药卫生—免疫学]