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Updated: Aug 12, 2026
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Expression of human single-chain variable fragment antibody against non-structural protein 3 of hepatitis C virus
1Gene Therapy Research Center, Institute of Infectious Diseases, 302nd Hospital of PLA, Beijing 100039, China.
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.
