title: [Construction and screening of hepatitis E virus-specific phage antibody combinatorial library].
abstract: OBJECTIVE
  To construct HEV-specific phage combinatorial anti-body library and screen anti-HEV antibodies with neutralizing activity from the library.
  
  
  METHODS
  The total RNA was extracted from B-lymphocytes of 6 HE patients. Kappa chain and Fd segment of IgG gene were amplified respectively by RT-PCR using a set of Fab-specific primers. The amplified gene were inserted successively into vector pComb3 and electrotransformed E. coli XLI-Blue cells. Furthermore, the recombinant phage was rescued by being concultured with helper phage VCSM13 to construct HEV-specific phage anti-body library.
  
  
  RESULTS
  Fab displayed on the surface a as fusion protein with the N terminal of coat protein III, and 1. 8 x 10(7) clone library was established. Specific antibodies to HEV ORF2 recombinant antigen were acquired after five rounds of panning with HEV ORF2 recombinant antigen including neutralizing epitope.
  
  
  CONCLUSIONS
  Four clones exhibited specific binding to HEV ORF2 recombinant antigen including neutralizing epitope is identified by ELISA. The results show that we have got the recombinant phage antibodies.
authors.affiliation: Genetic Research Center, Medical School, Southeast University, Nanjing 210009, China. wei.xie@seu.edu.cn
authors.forename: Hua
authors.initials: H
authors.lastname: Wei
authors.forename: Jian-Qiong
authors.initials: JQ
authors.lastname: Zhang
authors.forename: Hai-Qin
authors.initials: HQ
authors.lastname: Lu
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Meng
authors.forename: Xiao-Xuan
authors.initials: XX
authors.lastname: Lu
authors.forename: Wei
authors.initials: W
authors.lastname: Xie
mesh_terms: D006508:Hepatitis Antibodies / Q000235:genetics*; D016752:Hepatitis E virus / Q000276:immunology*; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D014764:Viral Proteins / Q000276:immunology*
chemical_list: D006508:Hepatitis Antibodies; D007145:Immunoglobulin kappa-Chains; C400971:ORF2 protein, Hepatitis E virus; D019151:Peptide Library; D014764:Viral Proteins