[Construction and expression of single chain Fv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum].
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.
Song
Xiao-tong
XT
Institute for Molecular Biology of Medicine, Nanjing Medical University, Nanjing 210029.
Feng
Zhen-qing
ZQ
Wang
Zhu-ming
ZM
Qiu
Zhen-ning
ZN
Li
Yun-qian
YQ
Guan
Xiao-hong
XH
Huang
Hua-liang
HL
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*
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