日本血吸虫抗独特型单克隆抗体NP30单链Fv基因的构建与表达

[Construction and expression of single chain Fv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum].

作者信息

Song Xiao-tong, Feng Zhen-qing, Wang Zhu-ming, Qiu Zhen-ning, Li Yun-qian, Guan Xiao-hong, Huang Hua-liang

机构信息

Institute for Molecular Biology of Medicine, Nanjing Medical University, Nanjing 210029.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2002;20(3):152-4.

PMID:12567991

Abstract

OBJECTIVE

To construct single chain Fv (scFv) gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum.

METHODS

The heavy and light chain variable region genes of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were inserted into two corresponding sites of expression vector pTHA90, and a scFv gene was constructed with a short peptide (Gly4Ser)3 linker gene. The recombinants were determined by digesting with XhoI/SpeI, XbaI/EcoRI and XhoI/EcoRI, and then were introduced into E. coli Top10. The antigen binding activity of expressed product was detected with ELISA.

RESULTS

The recombinants were determined by digesting with endonucleases and expected bands were identified. The value of expressed scFv was 3 times higher than negative control by ELISA(OD492 = 1.06).

CONCLUSION

The scFv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum was successfully cloned, and the expressed scFv fragment could interact specifically with antigen NP48.

摘要

目的

构建日本血吸虫抗独特型单克隆抗体NP30的单链Fv（scFv）基因。

方法

将日本血吸虫抗独特型单克隆抗体NP30的重链和轻链可变区基因插入表达载体pTHA90的两个相应位点，用短肽（Gly4Ser）3连接基因构建scFv基因。用XhoI/SpeI、XbaI/EcoRI和XhoI/EcoRI酶切鉴定重组体，然后导入大肠杆菌Top10。用ELISA检测表达产物的抗原结合活性。