中国寄生虫学与寄生虫病杂志 ›› 2004, Vol. 22 ›› Issue (2): 1-69.

• 论著 •    下一篇

溶组织内阿米巴重组人单克隆抗体Fab段的筛选与表达

舒勍1 ,邵红霞1,Hiroshi Tokaibana2,程训佳1   

  1. 1 复旦大学上海医学院病原生物系,上海200032 2 Tokai University School of Medicine,Japan
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2004-04-30 发布日期:2004-04-30

Screening and Expression of Recombinant Human Monoclonal Antibody Fab Fragments Specific to Entamoeba histolytica

SHU Qing,SHAO Hong-xia,Hiroshi Tachibana,CHENG Xun-jia   

  1. Department of Medical Microbiology & Parasitology,Shanghai Medical Center of Fudan University,Shanghai 200032,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2004-04-30 Published:2004-04-30

摘要: 目的 制备抗溶组织内阿米巴滋养体表面抗原的重组人单克隆抗体Fab段。 方法 提取无症状溶组织内阿米巴包囊携带者外周血淋巴细胞 ,分离总RNA ,行逆转录聚合酶链反应扩增IgG轻链和重链Fd片段 ,并与原核表达质粒连接 ,转化大肠埃希菌构建抗体库。通过抗原夹心膜法、间接荧光抗体试验等方法 ,筛选特异性表达抗溶组织内阿米巴滋养体表面抗原的重组人单克隆抗体Fab段的克隆 ,并纯化其表达产物。 结果 共筛选 3× 104个克隆 ,其中一个克隆表达的Fab段识别的抗原表位位于滋养体表面。 结论 运用原核表达系统可制备抗溶组织内阿米巴重组人单克隆抗体Fab段 ,在临床诊断和治疗上均有应用前景。

关键词: 溶组织内阿米巴, 单克隆抗体, 重组DNA, 基因表达

Abstract: Objective To prepare recombinant human monoclonal antibody Fab fragments specific to the surface antigen of Entamoeba histolytica. Methods Total RNA was isolated from lymphocytes which were separated from an asymptomatic E. histolytica cyst carrier. The genes of IgG light chain and Fd region of heavy chain were amplified by a reverse transcriptase PCR and ligated with a plasmid vector. After the genes were introduced into Escherichia coli, the clones expressing Fab fragments specific to the surface antigen of E. histolytica were screened and the product was purified. Results Thirty thousand clones were screened and one of them was proved positive to the surface antigen of E. histolytica. Conclusion This study demonstrated that the bacterial system can be used to produce recombinant human monoclonal antibody Fab fragments specific to the surface antigen of E. histolytica and they may be applicable for the future diagnosis and treatment of the infection.

Key words: Entamoeba histolytica, monoclonal antibodies, recombinant DNA, gene expression