Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli.
- Author:
Xiaobing XIA
1
;
Jun CHENG
;
Jizhen YANG
;
Yanwei ZHONG
;
Gang WANG
;
Hongqing FANG
;
Yan LIU
;
Ke LI
;
Jing DONG
Author Information
- Publication Type:Journal Article
- MeSH: Electrophoresis, Polyacrylamide Gel; Escherichia coli; genetics; Gene Expression; Hepatitis B Antibodies; biosynthesis; genetics; immunology; Hepatitis B Surface Antigens; immunology; Hepatitis B virus; drug effects; Humans; Immunoglobulin Fragments; biosynthesis; genetics; immunology; Interferon-alpha; biosynthesis; genetics; pharmacology; Recombinant Fusion Proteins; biosynthesis; genetics; pharmacology
- From: Chinese Journal of Hepatology 2002;10(1):28-30
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop a bacteria expression system to produce the fusion protein of humanized anti-HBsAg scFV and interferon-alpha.
METHODSThe expression vector was constructed after cleaving the plasmids harboring the humanized anti-HBsAg scFv and interferon alpha respectively and ligating to linearized pET22b subsequence. The expression of fusion protein in E.coli was analyzed by SDS-PAGE. The binding activity and antiviral activity of the fusion protein was characterized by competing inhibition test and cytopathic effect reduction.
RESULTSThe plasmid harboring the in frame arranged fusion gene was constructed and identified. After induction for 12h, a new band close to 4.5 10(4) was observed using SDS-PAGE. Results of competing ELISA and cytopathic effect reduction showed the fusion protein retained its specific binding activity and antiviral activities.
CONCLUSIONSThe construction and expression of the fusion gene of humanized anti-HBsAg scFv and interferon in E.coli are successful.
