Alphavirus capsid proteins self-assemble into core-like particles in insect cells: A promising platform for nanoparticle vaccine development
Autor(es): Hikke Mia C, Geertsema Corinne, Wu Vincen, Metz Stefan W, van Lent Jan W, Vlak Just M, Pijlman Gorben P
Resumo: The mosquito-borne chikungunya virus (CHIKV) causes arthritic diseases in humans, whereas the aquatic salmonid alphavirus (SAV) is associated with high mortality in aquaculture of salmon - trout. Using modern biotechnological approaches, promising vaccine c-idates based upon highly immunogenic, enveloped virus-like particles (eVLPs) have been developed. However, the eVLP structure (core, lipid membrane, surface glycoproteins) is more complex than that of non-enveloped, protein-only VLPs, which are structurally - morphologically 'simple'. In order to develop an alternative to alphavirus eVLPs, in this paper we engineered recombinant baculovirus vectors to produce high levels of alphavirus core-like particles (CLPs) in insect cells by expression of the CHIKV - SAV capsid proteins. The CLPs localize in dense nuclear bodies within the infected cell nucleus - are purified through a rapid - scalable protocol involving cell lysis, sonication - low-speed centrifugation steps. Furthermore, an immunogenic epitope from the alphavirus E2 glycoprotein can be successfully fused to the N-terminus of the capsid protein without disrupting the CLP self-assembling properties. We propose that immunogenic epitope-tagged alphavirus CLPs produced in insect cells present a simple - perhaps more stable alternative to alphavirus eVLPs.
Palavras-Chave: Alphavirus; Baculovirus expression; Core-like particle; Protein carrier; Subunit vaccine
Imprenta: Biotechnology Journal, v. 11, n. 2, p. 266-273, 2016
Identificador do objeto digital: 10.1002/biot.201500147
Descritores: Chikungunya virus - Cell ; Chikungunya virus - Proteins ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Vaccine
Data de publicação: 2016
