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Enterovirus71 virus‐like particles produced from insect cells and purified by multistep chromatography elicit strong humoral immune responses in mice (English)

In: Journal of applied microbiology   ;  119 ,  4  ;  2015
  • ISSN:
    1364-5072
  • Article (Journal)  /  Print

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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. 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. Our results demonstrated that EV71 VLPs can be purified with the multistep chromatographic protocol. 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.

Table of contents – Volume 119, Issue 4

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The tables of contents are generated automatically and are based on the data records of the individual contributions available in the index of the TIB portal. The display of the Tables of Contents may therefore be incomplete.

907
An overview on the origin and production of tetrodotoxin, a potent neurotoxin
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917
The use of probiotics in aquaculture
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Fate of Clostridium botulinum and incidence of pathogenic clostridia in biogas processes
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970
Differential expression of virulence genes and role of gyrA mutations in quinolone resistant and susceptible strains of Salmonella Weltevreden and Newport isolated from seafood
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Antimicrobial activity and chemometric modelling of South African propolis
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991
Efficacy of chemically characterized Foeniculum vulgare Mill seed essential oil in protection of raw tobacco leaves during storage against fungal and aflatoxin contamination
Kedia, A et al. | 2015
999
Pulsed light induced damages in Listeria innocua and Escherichia coli
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1011
Optimization of simultaneous removal of Cr (VI) and phenol by a native bacterial consortium: its use for bioaugmentation of co‐polluted effluents
Ontañon, O.M et al. | 2015
1023
Detection of biosurfactants in Bacillus species: genes and products identification
Płaza, G et al. | 2015
1035
Production and characterization of novel self‐assembling biosurfactants from Aspergillus flavus
Ishaq, U et al. | 2015
1046
Identification of chondroitin‐like molecules from biofilm isolates Exiguobacterium indicum A11 and Lysinibacillus sp. C13
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1057
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1064
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1075
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1101
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1127
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High prevalence of type 41 and high sequence diversity of partial hexon gene of human adenoviruses in municipal raw sewage and activated sludge
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1196
Enterovirus71 virus‐like particles produced from insect cells and purified by multistep chromatography elicit strong humoral immune responses in mice
Zhao, D et al. | 2015

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