title: [Construction and expression of humanized anti-HBsAg scFv targeting interferon-alpha in escherichia coli].
abstract: OBJECTIVE
  To develop a bacteria expression system to produce the fusion protein of humanized anti-HBsAg scFV and interferon-alpha.
  
  
  METHODS
  The 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The construction and expression of the fusion gene of humanized anti-HBsAg scFv and interferon in E.coli are successful.
authors.affiliation: Gene Therapy Research Centre, 302nd Hospital of PLA, Beijing 100039, China.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Xia
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.forename: Jizhen
authors.initials: J
authors.lastname: Yang
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Zhong
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Hongqing
authors.initials: H
authors.lastname: Fang
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Dong
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006510:Hepatitis B Antibodies / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006515:Hepatitis B virus / Q000187:drug effects; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology
chemical_list: D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007128:Immunoglobulin Fragments; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv