筛选出一株可识别多种变异HBsAg的单克隆抗体及其应用

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HBs Ag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.

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

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/c female mice were immunized with HBsAg isolated from serum of HBV infected patient.All monoclonal antibodies were produced according to standard methods.Total of 15 different HBsAg variants were produced in a mammalian 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(Hb1)was selected which could reacted strongly with most tested variants including G145R mutation.Conclusion The Hb1 is a valuable tool for detection of HBsAg variants.