1.Construction, expression and identification of a recombinant immunotoxin of scFv against Schistosoma japonicum
Hong LI ; Xiaojuan ZHU ; Chunyan GU ; Yongya REN ; Jing XU ; Zhongcan WANG ; Yuhua LI ; Zhenning QIU ; Jin ZHU ; Zhenqing FENG ; Xiaohong GUAN
Chinese Journal of Schistosomiasis Control 2010;22(2):117-121
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.
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