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.
Conclusion: 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.