title: Enterovirus71 virus-like particles produced from insect cells and purified by multistep chromatography elicit strong humoral immune responses in mice.
abstract: OBJECTIVE
  The study aims to develop a novel multistep chromatographic purification process for human enterovirus71 virus-like particles (VLPs) produced from insect cells (Sf9) infected with recombinant baculovirus.
  
  
  RESULTS
  Sf9 cells were maintained in the Wave Bioreactor system 20/50, and harvested when the viability decreased to 75% after infected with Bac-P1-3CD at the multiplicity of infection (MOI) of 1. After sonication and centrifugation, EV71 VLPs were purified with Capto(™) Core 700, Capto(™) adhere and Capto(™) butyl. The purity was then determined by SDS-PAGE, Western blotting and high-performance liquid chromatography (HPLC), while the diameter of purified EV71 VLPs was analysed by Dynamic Light Scattering (DLS) and Transmission electron microscopy (TEM). Immunization of BALB/c mice and serum collection were performed after contamination analysis, and neutralization antibodies were then analysed by pseudovirus-based microneutralization assay. Results showed that these purified EV71 VLPs can be successfully purified with ~31·52% yield and >95% purity. They could elicit stronger neutralization antibodies in mice compared with those produced from formalin-inactivated EV71 virus.
  
  
  CONCLUSIONS
  Our results demonstrated that EV71 VLPs can be purified with the multistep chromatographic protocol.
  
  
  CONCLUSIONS
  This work presents a novel multistep chromatographic technique, an effective way of purifying EV71 VLPs with high purity. This purification process can thus serve as foundation for further development of industrial-scale production process of EV71 VLP vaccine candidates.
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: D
authors.initials: D
authors.lastname: Zhao
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.|Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, Jilin University, Changchun, China.
authors.forename: B
authors.initials: B
authors.lastname: Sun
authors.affiliation: The China-Japan Fellowship Hospital of Jilin University, Changchun, China.
authors.forename: H
authors.initials: H
authors.lastname: Jiang
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: S
authors.initials: S
authors.lastname: Sun
authors.affiliation: Ann Arbor Pioneer High School, Ann Arbor, MI, USA.
authors.forename: F T
authors.initials: FT
authors.lastname: Kong
authors.affiliation: Changchun BCHT Biotechnology Company, Changchun, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Ma
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: L
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Changchun BCHT Biotechnology Company, Changchun, China.
authors.forename: L
authors.initials: L
authors.lastname: Bai
authors.affiliation: Changchun BCHT Biotechnology Company, Changchun, China.
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.affiliation: Changchun BCHT Biotechnology Company, Changchun, China.
authors.forename: P
authors.initials: P
authors.lastname: Yang
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: C
authors.initials: C
authors.lastname: Liu
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Xu
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.|Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, Jilin University, Changchun, China.|State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, China.
authors.forename: W
authors.initials: W
authors.lastname: Su
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.|Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, Jilin University, Changchun, China.
authors.forename: W
authors.initials: W
authors.lastname: Kong
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.|Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, Jilin University, Changchun, China.
authors.forename: F
authors.initials: F
authors.lastname: Xu
authors.affiliation: School of Life Sciences, Jilin University, Changchun, China.|National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, China.|Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, Jilin University, Changchun, China.
authors.forename: C
authors.initials: C
authors.lastname: Jiang
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D002460:Cell Line; D002845:Chromatography / Q000379:methods*; D029821:Enterovirus A, Human / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004769:Enterovirus Infections / Q000276:immunology* / Q000821:virology; D005260:Female; D006801:Humans; D056724:Immunity, Humoral; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D018411:Spodoptera
chemical_list: D057134:Antibodies, Neutralizing
keywords: chromatography; enterovirus 71; immunogenicity; insect cells; virus-like particle