Enterovirus71 virus-like particles produced from insect cells and purified by multistep chromatography elicit strong humoral immune responses in mice.

Abstract

AIMS:

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.

METHODS AND 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.

SIGNIFICANCE AND IMPACT OF THE STUDY:

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: D Zhao, B Sun, H Jiang, S Sun, F T Kong, Y Ma, L Jiang, L Bai, X Chen, P Yang, C Liu, Y Xu, W Su, W Kong, F Xu, C Jiang
• Journal: Journal of applied microbiology (J Appl Microbiol) Vol. 119 Issue 4 Pg. 1196-205 (Oct 2015) ISSN: 1365-2672 [Electronic] England
• PMID: 26248815 (Publication Type: Evaluation Study, Journal Article)
• Copyright: © 2015 The Society for Applied Microbiology.

Chemical References

• Antibodies, Neutralizing

Topics

• Animals
• Antibodies, Neutralizing (immunology)
• Cell Line
• Chromatography (methods)
• Enterovirus A, Human (chemistry, genetics, immunology, isolation & purification)
• Enterovirus Infections (immunology, virology)
• Female
• Humans
• Immunity, Humoral
• Immunization
• Mice
• Mice, Inbred BALB C
• Molecular Sequence Data
• Spodoptera