title: [Purification and immunological characterization of hepatitis E virus recombinant chimeric antigen encoded by ORF2 fragments and ORF3].
abstract: OBJECTIVE
  To study immunological characteristics of recombinant chimeric HEV antigen.
  
  
  METHODS
  Constructed recombinant plasmids pThioHisORF(2.1 + 2.2 + 3), which contains three HEV antigen gene fragments (ORF2.1:6287-6403nt, ORF2.2:6743-7126nt, ORF3), was transformed into E. coli and induced with IPTG. Expressed product P(2.1 + 2.2 + 3) existed in inclusion bodies, was purified by denature SP Sepharose FF cation exchange chromatography. Rabbits and rats were immunized with renatured P(2.1 + 2.2 + 3). The level of IgG in sera from experimental animals and clinical patients were examined with P(2.1 + 2.2 + 3) by ELISA. The characteristics of IgG of immunized animals interacted with recombinant antigen expressed by baculovirus system as well as recombinant chimeric antigen interacted with clinical patients sera were evaluated by Western-blotting.
  
  
  RESULTS
  High titer of IgG antibodies, 1:25,600 in rabbits and 1:12,800 in rats, were detected after immunized with P(2.1 + 2.2 + 3). Furthermore, recombinant antigen expressed by baculovirus system was specifically recognized by IgG of experimental animal immunized with P(2.1 + 2.2 + 3), and the purified recombinant chimeric antigen P(2.1 + 2.2 + 3) was specifically reacted with the IgG of clinical patients.
  
  
  CONCLUSIONS
  Recombinant chimeric antigen appears a promising strategy for detection of and prevention from HEV infection.
authors.affiliation: Department of Molecular Biology, Institute of Medical Biology, CAMS and PUMC, Kunming 650118, China.
authors.forename: H
authors.initials: H
authors.lastname: Tang
authors.forename: Y B
authors.initials: YB
authors.lastname: Ma
authors.forename: R J
authors.initials: RJ
authors.lastname: Du
authors.forename: G Y
authors.initials: GY
authors.lastname: Le
authors.forename: C H
authors.initials: CH
authors.lastname: Li
authors.forename: J Y
authors.initials: JY
authors.lastname: Zhuang
authors.forename: Y
authors.initials: Y
authors.lastname: Liu
authors.forename: M S
authors.initials: MS
authors.lastname: Sun
authors.forename: C B
authors.initials: CB
authors.lastname: Dai
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D016752:Hepatitis E virus / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification*; D014764:Viral Proteins / Q000276:immunology* / Q000302:isolation & purification*
chemical_list: C400971:ORF2 protein, Hepatitis E virus; C400973:ORF3 protein, Hepatitis E virus; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins