Production Of Recombinant Virus-like Particles Research Articles

High-yield production of recombinant virus-like particles of enterovirus 71 in Pichia pastoris and their protective efficacy against oral viral challenge in mice

Enterovirus 71 (EV71) is one of the major causative pathogens of hand, foot and mouth disease (HFMD), which is highly prevalent in the Asia-Pacific regions. Severe HFMD cases with neurological complications and even death are often associated with EV71 infections. However, no licensed EV71 vaccine is currently available. Recombinant virus-like particles (VLPs) of EV71 have been produced and shown to be a promising vaccine candidate in preclinical studies. However, the performance of current recombinant expression systems for EV71 VLP production remains unsatisfactory with regard to VLP yield and manufacturing procedure, and thus hinders further product development. In this study, we evaluated the expression of EV71 VLPs in Pichia pastoris and determined their protective efficacy in mouse models of EV71 infections. We showed that EV71 VLPs could be produced at high levels up to 4.9% of total soluble protein in transgenic P. pastoris yeast co-expressing P1 and 3CD proteins of EV71. The resulting yeast-produced VLPs potently induced neutralizing antibodies against homologous and heterologous EV71 strains in mice. More importantly, maternal immunization with VLPs protected neonatal mice in both intraperitoneal and oral challenge experiments. Collectively, these results demonstrated the success of simple, high-yield production of EV71 VLPs in transgenic P. pastoris, thus lifting the major roadblock in commercial development of VLP-based EV71 vaccines.

Virus-Like Particles Expressing the Nucleocapsid Gene as an Efficient Vaccine Against Rift Valley Fever Virus

Rift Valley fever virus (RVFV), a member of the family Bunyaviridae, regularly accounts for large and severe outbreaks among humans and livestock in Africa and Arabia. Therefore, safe and efficient vaccines are highly needed. Here, we report the production of recombinant virus-like particles (VLPs) that, in addition to their similarity to RVFV particles, are able to express the viral nucleocapsid (N) gene. A single inoculation of 1 × 10(6) of these N-VLPs was sufficient to protect 100% of mice from infection with a lethal dose of 1 × 10(5) PFU of RVFV. Our study demonstrates that N-VLPs can be considered as a safe and efficient vaccine against the emerging pathogen RVFV, and that VLPs that actively produce a viral antigen may be considered a strategy to improve the immunogenicity of VLPs in general.

Production of recombinant virus-like particles from human papillomavirus types 6 and 11, and study of serological reactivities between HPV 6, 11, 16 and 45 by ELISA: implications for papillomavirus prevention and detection

The L1 major capsid proteins of human papillomaviruses types 6 and 11 were expressed in insect cells using recombinant baculoviruses. These L1 proteins were shown to self-assemble into virus-like particles resembling papillomavirus virions as previously observed for HPV 16 and 45. However, we observed variations in the yield of virus-like particles among the four genotypes investigated. This suggests that more than one strain of each genotype has to be investigated to obtain the high level of virus-like particle production necessary to develop HPV vaccines or serological tests. Cross-reactivities between HPV 6, 11, 16 and 45 were studied using polyclonal and monoclonal antibodies to virus-like particles, L1 proteins and synthetic peptides. Although antisera react strongly against homologous virus-like particles, there is evidence of some cross-reactivity. This could be one of the explanations for the fact that antibodies to one genotype are detected in individuals infected with another genotype. This study also identified a linear epitope recognized by anti-HPV 16 virus-like particle sera.

Production of recombinant virus-like particles from human papillomavirus types 6 and 11, and study of serological reactivities between HPV 6, 11, 16 and 45 by ELISA: implications for papillomavirus prevention and detection.

The L1 major capsid proteins of human papillomaviruses types 6 and 11 were expressed in insect cells using recombinant baculoviruses. These L1 proteins were shown to self-assemble into virus-like particles resembling papillomavirus virions as previously observed for HPV 16 and 45. However, we observed variations in the yield of virus-like particles among the four genotypes investigated. This suggests that more than one strain of each genotype has to be investigated to obtain the high level of virus-like particle production necessary to develop HPV vaccines or serological tests. Cross-reactivities between HPV 6, 11, 16 and 45 were studied using polyclonal and monoclonal antibodies to virus-like particles, L1 proteins and synthetic peptides. Although antisera react strongly against homologous virus-like particles, there is evidence of some cross-reactivity. This could be one of the explanations for the fact that antibodies to one genotype are detected in individuals infected with another genotype. This study also identified a linear epitope recognized by anti-HPV 16 virus-like particle sera.

Spherical virus assembly

A wide variety of structural and biophysical techniques have been used to study viral assembly pathways. For example, the use of three-dimensional electron microscopy has allowed the visualization of viral assembly intermediates, as well as mature capsids. More detailed views of small icosahedral viruses and viral proteins, such as proteases, have been provided by X-ray crystallography. The combination of these structural techniques represents a powerful new approach to investigating large assemblies in atomic detail. The production of recombinant virus-like particles has become another popular approach to studying viral assembly.