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

@entity1 @entity9010 @entity19 @entity406 @entity281
AIMS: The study aims to develop a novel multistep chromatographic purification process for @entity1 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 @entity281 (MOI) of 1. After sonication and centrifugation, EV71 VLPs were purified with Capto(   ) Core 700, Capto(   ) adhere and Capto(   ) @entity9010 . 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 @entity19 compared with those produced from @entity406 -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.
