Expression of human single-chain variable fragment antibody against non-structural protein 3 of hepatitis C virus antigen in e.coli.
OBJECTIVE
To express human single-chain variable fragment (ScFv) antibody against non-structural protein 3 (NS(3)) of hepatitis C virus in E.coli.


METHODS
The recombinant phages were panned by NS(3) antigen which was coated in a microtiter plate. After five rounds of biopanning, 66 clones were identified specific to NS(3) antigen. E.coli host XL(1)-Blue was transformed and induced by IPTG. The specificity of ScFv was evaluated by ELISA and dot blot hybridization.


RESULTS
ScFv-NS(3) DNA digestion and sequencing data showed that the ScFv gene was composed of 750bp. ELISA and dot blot hybridization demonstrated that the soluble human single-chain Fv antibody to hepatitis C virus NS(3) antigen could combine different origins of NS(3) antigen.


CONCLUSIONS
NS(3)-ScFv antibody expressed by E.coli host XL(1)-Blue has the activity and specificity to combine different origins of HCV NS(3) antigen.
Zhong
Y
Y
Gene Therapy Research Center, Institute of Infectious Diseases, 302nd Hospital of PLA, Beijing 100039, China.
Cheng
J
J
Liu
Y
Y
Dong
J
J
Yang
J
J
Zhang
L
L
D004926:Escherichia coli / Q000235:genetics; D018937:Hepatitis C Antibodies / Q000096:biosynthesis*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D018937:Hepatitis C Antibodies; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C055966:immunoglobulin Fv