Construction, expression and identification of a recombinant immunotoxin of scFv against Schistosoma japonicum
- VernacularTitle:抗日本血吸虫单链抗体免疫毒素的构建表达及生物学活性鉴定
- Author:
Hong LI
;
Xiaojuan ZHU
;
Chunyan GU
;
Yongya REN
;
Jing XU
;
Zhongcan WANG
;
Yuhua LI
;
Zhenning QIU
;
Jin ZHU
;
Zhenqing FENG
;
Xiaohong GUAN
- Publication Type:Journal Article
- Keywords:
Schistosoma japonicum;
scFv;
PE38KDEL;
Recombinant immunotoxin
- From:
Chinese Journal of Schistosomiasis Control
2010;22(2):117-121
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a recombinant immunotoxin expression vector composed of a single-chain Fv fragment of Sehistosorna japomicum and PE38KDEL gene,and identify the binding activity of the purified product with SEA antigen.Methods The V_H and V_L genes were amplified by PCR from the parent monoclonal antibody NP11-4.Then the amplified scFv and PE38KDEL genes were inserted into the expression vector pBAD/gIII A.The fusion protein expressed in E.coli Top10F' induced by L-arabinose.After purification,the activity of the immunotoxin was evaluated by Westem-blot and ELISA.Results The new recombinant immunotoxin expression vector pBAD/gIII A-scfv-PE38KDEL was constructed successfully.The main product was in inclusion bodies.ELISA assay showed that the refolding recombinant immunotoxin remained binding activity with SEA antigen.Conclusion A new recombinant expression plasmid pBAD/gIII A-scfv-PE38KDEL has been constructed and expressed successfully,which is useful in further study of the treatment of schistosomiasis japonica.
