title: [Expression of structural protein VP1 of enterovirus 71 in E.coli and preparation of anti-VP1 monoclonal antibodies].
abstract: OBJECTIVE
  To express recombinant structural protein VP1 of enterovirus 71 (EV71) in E.coli and prepare anti-VP1 monoclonal antibodies (mAbs).
  
  
  METHODS
  With P1 gene as a template,    EV71 VP1 gene was amplified by PCR and cloned into the expression vector pET-32a(+). After transformation into E.coli TG1,    the positive clones were screened and sequenced. The recombinant plasmid was then transformed into BLgold (DE3) and the recombinant protein in inclusion bodies was induced by IPTG and detected by SDS-PAGE. After the inclusion bodies were solubilized with 6 mol/L guanidine hydrochloride,    we purified VP1 protein using Ni-NTA affinity chromatography,    and then immunized the mice with it to prepare the mAbs against VP1 protein.
  
  
  RESULTS
  Recombinant VP1 protein was expressed in E.coli. We obtained totally 24 VP1 monoclonal hybridoma cell strains,    of which one  was EV71  positive determined by Western blotting  and five were  positive for IFA.
  
  
  CONCLUSIONS
  These mAbs are valuable reagents for the  development  of new vaccines and detection kits for EV71.
authors.affiliation: Guangzhou Ridabiotech Co., LTD, Guangzhou, China.
authors.forename: Chen-yang
authors.initials: CY
authors.lastname: Li
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.forename: Yan-li
authors.initials: YL
authors.lastname: Gu
authors.forename: Ya-nan
authors.initials: YN
authors.lastname: Gu
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Chen
authors.forename: Lu
authors.initials: L
authors.lastname: Tian
authors.forename: Li-yun
authors.initials: LY
authors.lastname: Chen
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D029821:Enterovirus A, Human / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015678:Viral Structural Proteins / Q000096:biosynthesis* / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D015678:Viral Structural Proteins; D014765:Viral Vaccines