抗HBV preS1嵌合抗体的真核表达及活性分析  

Expression and activity analysis of chimeric antibody against EIBV preS1

作　　者：陈瑛炜[1] 罗文新[1] 王晋[1] 王明桥[1] 顾颖[1] 林亦伟[1] 张军[1] 夏宁邵[1] 

机构地区：[1]国家传染病诊断试剂与疫苗工程技术研究中心,厦门大学细胞生物学与肿瘤细胞工程教育部重点实验室,厦门大学生命科学学院,361005

出　　处：《中华微生物学和免疫学杂志》2007年第2期167-171,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（No.30640017）;国家十五创新药物博士基金（No.2003AA223539）;厦门市科技计划项目（No.350Z220055002）

摘　　要：目的 通过真核系统表达抗HBVpreS1嵌合抗体4D11并分析其活性。方法 通过RT-PCR方法从分泌抗乙型肝炎病毒preS1的鼠源单克隆抗体4D11的杂交瘤细胞中克隆出抗体基因的重链可变区（VH）、轻链可变区（Vk）序列，并分别克隆到含有人71重链和X轻链恒定区序列的pcDNA3．1-AH和pcDNA3．1-Ak质粒中，共转染人胚肾细胞变种（293FT）细胞。通过Western blot、竞争ELISA、阳性血清阻断及血清中preS1的检测实验来分析其活性。结果 轻重链基因在细胞内正确组装成嵌合抗体分子并分泌至细胞外，具有结合活性。嵌合抗体可以很好的与病毒preSl结合，对阳性血清的检测效果与鼠单抗一致。结论 成功的利用293FT细胞表达了4D11嵌合抗体，抗体保留了原鼠单抗的特异性和活性，为探讨乙肝抗体的治疗提供了试验资料。Objective To express the chimeric anti-HBV preS1 antibody 4D11 in eukaryotic system and analyze the activity. Methods cDNAs encoding variable regions of heavy and fight chains were isolated by RT-PCR from hybridoma cells secreting mouse monoclonal antibody （McAb） 4D11, and to be introduced into mam- malian expression vectors pcDNA3.1-AH and pcDNA3.1-Ak containing sequences of human T1 and g constant regions, respectively. The vectors were cotransfected into 293 FT cells. The expressed 4D11 chimeric antibody （cAb） was characterized by Western blot, competitive ELISA, serum blocking ELISA and preS1 detection insenan. Results The expressed heavy and light chains in 293FF cells were assembled to form IgG molecule, which could be secreted outside the cells. The cAb well reacted with preS1 in HBV eAg positive serum. The result of preS1 detection in HBV eAg positive serum by gl,l.qA, with coated cAb was consistent with that of4Dll McAb. Conclusion cAb expressed in 293 Fr cells, shows the same binding specificity and recongnizes the same epitope as original McAb. The cAb described here is expected to be less immunoreactive and thus more suitable for antibody therapy of hepatitis B.

关 键 词：乙型肝炎病毒 嵌合抗体 真核表达 

分 类 号：R686[医药卫生—骨科学]