title: Characterizing Enterovirus 71 and Coxsackievirus A16 virus-like particles production in insect cells.
abstract: Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are two viruses commonly responsible for hand, foot and mouth disease (HFMD) in children. The lack of prophylactic or therapeutic measures against HFMD is a major public health concern. Insect cell-based EV71 and CVA16 virus-like particles (VLPs) are promising vaccine candidates against HFMD and are currently under development. In this paper, the influence of insect cell line, incubation temperature, and serial passaging effect and stability of budded virus (BV) stocks on EV71 and CVA16 VLP production was investigated. Enhanced EV71 and CVA16 VLP production was observed in Sf9 cells compared to High Five™ cells. Lowering the incubation temperature from the standard 27°C to 21°C increased the production of both VLPs in Sf9 cells. Serial passaging of CVA16 BV stocks in cell culture had a detrimental effect on the productivity of the structural proteins and the effect was observed with only 5 passages of BV stocks. A 2.7× higher production yield was achieved with EV71 compared to CVA16. High-resolution asymmetric flow field-flow fractionation couple with multi-angle light scattering (AF4-MALS) was used for the first time to characterize EV71 and CVA16 VLPs, displaying an average root mean square radius of 15±1nm and 15.3±5.8 nm respectively. This study highlights the need for different approaches in the design of production process to develop a bivalent EV71 and CVA16 vaccine.
authors.affiliation: The University of Queensland, Protein Expression Facility, Brisbane, QLD 4072, Australia.
authors.forename: Balaji
authors.initials: B
authors.lastname: Somasundaram
authors.affiliation: The University of Queensland, Protein Expression Facility, Brisbane, QLD 4072, Australia.
authors.forename: Cindy
authors.initials: C
authors.lastname: Chang
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD 4072, Australia.
authors.forename: Yuan Y
authors.initials: YY
authors.lastname: Fan
authors.affiliation: Sentinext Therapeutics Sdn Bhd, Sains@USM, 10050 Penang, Malaysia.
authors.forename: Pei-Yin
authors.initials: PY
authors.lastname: Lim
authors.affiliation: Sentinext Therapeutics Sdn Bhd, Sains@USM, 10050 Penang, Malaysia.
authors.forename: Jane
authors.initials: J
authors.lastname: Cardosa
authors.affiliation: The University of Queensland, Protein Expression Facility, Brisbane, QLD 4072, Australia. Electronic address: l.lua@uq.edu.au.
authors.forename: Linda
authors.initials: L
authors.lastname: Lua
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D004770:Enterovirus / Q000235:genetics / Q000276:immunology*; D029821:Enterovirus A, Human / Q000235:genetics / Q000276:immunology*; D006232:Hand, Foot and Mouth Disease / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D006801:Humans; D018716:Molecular Mimicry; D011817:Rabbits; D061987:Sf9 Cells; D018411:Spodoptera; D013696:Temperature; D058425:Vaccines, Virus-Like Particle / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014771:Virion / Q000235:genetics* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D058425:Vaccines, Virus-Like Particle; D014765:Viral Vaccines
keywords: Enterovirus; Field-flow fractionation; Insect cell; Vaccine; Virus-like particle