Formation of virus-like particles from O-type foot-and-mouth disease virus in insect cells using codon-optimized synthetic genes

Abstract

A recombinant baculovirus was constructed to simultaneously express codon-optimized virus-like particles (VLP), A VP1-2A-VP3 and VP0 of serotype O foot-and-mouth disease virus (FMDV), from individual promoters. The target proteins were expressed in insect cells at high level, as shown by indirect sandwich ELISA; and the expressed VP1-2A-VP3 could autocatalytically be cleaved into the individual proteins, VP1-2A and VP3, as shown by Western-blot analyses. In addition, in the insect cells, the structural proteins, VP0, VP3 and VP1-2A, self-assembled into virus-like particles resembling the authentic FMDV particles. This information should prove useful for the development of more efficient VLP assembly using shorter genes.

Foot-and-mouth disease virus-like particles produced by a SUMO fusion protein system in Escherichia coli induce potent protective immune responses in guinea pigs, swine and cattle

“…These proteins can then self-assemble to form icosahedral empty capsid particles [31-33], which consist of 60 copies of each protein. Many attempts have been made to produce FMDV capsid protein(s) and subsequently assemble into VLP in eukaryotic expression systems by combining the P1-2A and 3C, in order to assemble virus capsids correctly [12,34]. However, this strategy has not been achieved in prokaryotic expression systems, especially in E. coli .…”

Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity

“…Similarly, a dual expression strategy of an Asia 1 serotype of FMDV led to incomplete cleavage of the P1-2A precursor (Cao et al, 2009). More recently, forsaking the use of 3C to generate the mature capsid proteins, VP0 and VP3-2A-VP1 from an O serotype of FMDV were co-expressed relying on self-cleavage at the 2A site (Donnelly et al, 2001) to generate the requisite structural proteins for assembly, which resulted in partial success (Cao et al, 2010). In a further example, a Bombyx (silk worm) baculovirus system encoding a P1-2A-3C sequence of an FMDV Asia 1 isolate was used as a single transcription unit driven by the polyhedrin promoter and the empty capsid material harvested from the haemolymph of the infected silk worms was immunogenic in cattle and led to levels of neutralising antibody associated with protection (Li et al, 2008, 2011).…”

Specific detection of foot-and-mouth disease serotype Asia 1 virus by carboxyl-magnetic beads conjugated with single-domain antibody

“…[ 21 ]. The recombinant FMDV VLP contains three structural proteins, VP0, VP1 and VP3 in vivo when observed by SDS-PAGE [ 22 ]. We calculated that 1.0 μg of MNB (approximately 1.2 × 10 8 beads) are able to bind 5.09 μg target antigen.…”