Development of EV71 virus-like particle purification processes
Journal
Vaccine
Journal Volume
33
Journal Issue
44
Pages
5966-5973
Date Issued
2015
Author(s)
Abstract
Enterovirus 71 (EV71) causes the outbreaks of hand-foot-and-mouth disease and results in deaths of hundreds of young children. EV71 virus-like particles (VLPs) are empty capsids consisting of viral structural proteins and can elicit potent immune responses, thus holding promise as an EV71 vaccine candidate. However, an efficient, scalable production and purification scheme is missing. For mass production of EV71 VLPs, this study aimed to develop a production and chromatography-based purification process. We first demonstrated the successful EV71 VLPs production in the stirred-tank bioreactor in which High Five? cells were infected with a recombinant baculovirus co-expressing EV71 structural polyprotein P1 and protease 3CD. The culture supernatant containing the VLPs was subjected to tangential flow filtration (TFF) for concentration/diafiltration, which enabled the removal of >80% of proteins while recovering >80% of VLPs. The concentrated VLPs were next subjected to hydroxyapatite chromatography (HAC) in which the VLPs were mainly found in the flow through. After another TFF concentration/diafiltration, the VLPs were purified by size-exclusion chromatography (SEC) and concentrated/diafiltered by a final TFF. The integrated process yielded an overall VLPs recovery of ?36% and a purity of ?83%, which was better or comparable to the recovery and purity for the purification of live EV71 virus particles. This process thus may move the EV71 VLPs vaccine one step closer to the clinical applications. ? 2015 Elsevier Ltd.
Subjects
Chromatography; Downstream processing; Enterovirus 71; Purification; Vaccine; Virus-like particle
SDGs
Other Subjects
virus protein; recombinant protein; virosome; virus like particle vaccine; anion exchange; antigenicity; Article; cell viability; cytolysis; diafiltration; Enterovirus 71; immune response; isoelectric point; large scale production; molecular weight; nonhuman; priority journal; size exclusion chromatography; vaccine production; virus like agent; virus particle; virus purification; animal; Baculoviridae; bioreactor; cell line; Enterovirus A; filtration; genetics; growth, development and aging; insect; isolation and purification; liquid chromatography; procedures; Animals; Baculoviridae; Bioreactors; Cell Line; Chromatography, Gel; Chromatography, Liquid; Enterovirus A, Human; Filtration; Insects; Recombinant Proteins; Vaccines, Virus-Like Particle; Virosomes
Publisher
Elsevier Ltd
Type
journal article