Y Zhong1, J Cheng, Y Liu, J Dong, J Yang, L Zhang. 1. Gene Therapy Research Center, Institute of Infectious Diseases, 302nd Hospital of PLA, Beijing 100039, China.
Abstract
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.
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.
Authors: Tian-Cheng Li; Naokazu Takeda; Tatsuo Miyamura; Yoshiharu Matsuura; Joseph C Y Wang; Helena Engvall; Lena Hammar; Li Xing; R Holland Cheng Journal: J Virol Date: 2005-10 Impact factor: 5.103