title: [Construction and expression of single chain Fv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum].
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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute for Molecular Biology of Medicine, Nanjing Medical University, Nanjing 210029.
authors.forename: Xiao-tong
authors.initials: XT
authors.lastname: Song
authors.forename: Zhen-qing
authors.initials: ZQ
authors.lastname: Feng
authors.forename: Zhu-ming
authors.initials: ZM
authors.lastname: Wang
authors.forename: Zhen-ning
authors.initials: ZN
authors.lastname: Qiu
authors.forename: Yun-qian
authors.initials: YQ
authors.lastname: Li
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Guan
authors.forename: Hua-liang
authors.initials: HL
authors.lastname: Huang
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis / Q000235:genetics*; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000235:genetics*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D007128:Immunoglobulin Fragments / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D012549:Schistosoma japonicum / Q000276:immunology*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; C055966:immunoglobulin Fv