南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (02): 253-.

• • 上一篇    下一篇

HBeAg的B细胞线性表位预测及鉴定

杨军，刘妮，张婷，赵世平，强磊，苏宝山，康安静，李宗芳   

  • 出版日期:2013-02-20 发布日期:2013-02-20

Prediction and identification of B-cell linear epitopes of hepatitis B e antigen

  • Online:2013-02-20 Published:2013-02-20

PDF

2071

摘要： 目的预测并鉴定乙型肝炎病毒e抗原（HBeAg）的B细胞线性表位，为乙型肝炎的诊断和治疗提供新的依据。方法采用
生物信息学分析技术，利用NCBI数据库和免疫表位数据库提供的相应软件预测HBeAg的B细胞线性表位，采用人工合成法合
成相应表位肽并分别将与血蓝蛋白（KLH）偶联，作为免疫原，免疫大白兔制备抗HBeAg抗原表位抗体，ELISA法鉴定抗体的特
异性。结果发现了1MDIDPYKEFG10、37LYREALESPEHCSP50、74SNLEDPAS81、127RTPPAYRPPNAPIL140等4条新的HBeAg蛋白
B细胞线性表位肽，其与KLH的偶联物作为免疫原免疫大白兔，获得特异性高效价抗体，抗体滴度大于1∶512000，ELISA实验
证实上述抗体均可与HBeAg发生特异性免疫反应。结论采用生物信息学技术成功确认了4个HBeAg蛋白B细胞线性表位
肽，为深入研究HBeAg的功能和作用以及乙型肝炎的治疗提供了新依据。

Abstract: Objective To predict and identify B-cell linear epitopes of hepatitis B e antigen (HBeAg). Methods The B-cell linear
epitopes of HBeAg were predicted using the software provided by NCBI Database and Immune Epitope Database (IEDB) and
synthesized by a solid-phase method followed by conjugation with keyhole limpet hemocyanin (KLH). The KLH conjugates
were used for immunization of New Zealand white rabbits, and the immune response of the rabbits was monitored by direct
ELISA using a bovine serum albumin conjugate of the predicted epitopes. Results Four new B-cell linear epitopes of HBeAg
were identified, namely 1MDIDPYKEFG10, 37LYREALESPEHCSP50, 74SNLEDPAS81 and 127RTPPAYRPPNAPIL140. The rabbits
immunized with the KLH conjugate showed an antibody titer over 1∶512 000. The antisera of B-cell linear epitopes collected
could specifically react with HBeAg as shown by ELISA. Conclusion Four B-cell linear epitopes of HBeAg have been
confirmed using bioinformatics methods, which provides new evidence for further functional studies of HBeAg in hepatitis B.