Construction of polyomavirus-derived pseudotype virus-like particles displaying a functionally active neutralizing antibody against hepatitis B virus surface antigen

Plečkaitytė, Milda; Bremer, Corinna M.; Gedvilaitė, Alma; Kučinskaitė-Kodzė, Indrė; Glebe, Dieter; Žvirblienė, Aurelija

doi:10.1186/s12896-015-0203-3

Cited by 22 publications

(21 citation statements)

References 37 publications

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“…Moreover, hamster polyomavirus VLPs comprising native VP1 and chimeric VP2 harbouring an anti‐HBsAg antibody molecule have been produced in S. cerevisiae , and they allowed surface exposure of functionally active neutralizing antibody against HBsAgs (Pleckaityte et al . ).…”

Section: Yeast‐derived Vlps As Delivery Systems For Antigens Nucleicmentioning

confidence: 97%

Yeast as an expression system for producing virus-like particles: what factors do we need to consider?

Kim

2016

Lett Appl Microbiol

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The development of various types of virus-like particles (VLPs) has accelerated over the past two decades as the importance of VLPs for generating nextgeneration vaccines has been appreciated. Yeast has advantages such as scalable fermentation, low risk of contamination by adventitious agents, low production costs and the ability to produce VLPs with reliable qualities. It is generally recognized that yeast is suitable for producing VLPs that have simple structures and are produced intracellularly. However, recently there has been much effort to extend its applicability, and there is now evidence that it can be used as an expression platform for the productions of VLPs not only of nonenveloped viruses but also of enveloped viruses. Moreover, evidences indicated that yeast allows secretory VLP productions. Meanwhile, it has become evident that the quality and quantity of yeast-derived VLPs are influenced by the choice of plasmid and promoter, the ratio of the structural proteins produced. Here, we review the characteristics of the yeast expression system in terms of the production of VLP and compare it with other expression systems. We also consider strategies for VLP production in yeast and factors that need to be taken into account.

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Section: Yeast‐derived Vlps As Delivery Systems For Antigens Nucleicmentioning

confidence: 97%

Yeast as an expression system for producing virus-like particles: what factors do we need to consider?

Kim

2016

Lett Appl Microbiol

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“…There exists another approach to designing therapeutic vaccines based on VLPs. In this case, the VLP surface displays the variable fragment of an antibody specific to the antigen of the target virus [65]. Moreover, knowing which receptor is bound by the virus proteins renders it possible to produce particles possessing tropism to the cells of certain tissues.…”

Section: Recombinant Vlps For Presentation Of Foreign Antigenic Determentioning

confidence: 99%

Virus-Like Particles as an Instrument of Vaccine Production

Syomin

Ilyin

2019

Mol Biol

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The paper discusses the techniques which are currently implemented for vaccine production based on virus-like particles (VLPs). The factors which determine the characteristics of VLP monomers assembly are provided in detail. Analysis of the literature demonstrates that the development of the techniques of VLP production and immobilization of target antigens on their surface have led to the development of universal platforms which make it possible for virtually any known antigen to be exposed on the particle surface in a highly concentrated form. As a result, the focus of attention has shifted from the approaches to VLP production to the development of a precise interface between the organism's immune system and the peptides inducing a strong immune response to pathogens or the organism's own pathological cells. Immunome-specified methods for vaccine design and the prospects of immunoprophylaxis are discussed. Certain examples of vaccines against viral diseases and cancers are considered.

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“…GFP was fused to the C-terminus of VP2C and was tagged with 6His to enable purification. Unlike previous reports where GFP was fused to the N-terminus of MPyV VP2 (Abbing et al 2004;Boura et al 2005), in my research GFP was fused to C-terminus of VP2C based on the findings of Inoue et al (2008) with SV40 andPleckaityte et al (2015) with HPyV, that suggested a complete encapsidation with C-terminal fusion.…”

Section: Optimisation Of Encapsidationmentioning

confidence: 90%

“…Fusion at the C-terminus of VP2C is expected to position GFP inside the cavity of the VLP (Inoue et al 2008;Pleckaityte et al 2015). This attempt was done by two approaches, the first is to bind VP2C-GFP and VP1 capsomeres in vitro, and the second is to facilitate binding of VP2C-GFP and VP1 capsomeres in vivo.…”

Section: Resultsmentioning

confidence: 99%

“…Fusion of GFP protein at the N-terminus of VP2 was attempted to encapsidate GFP inside the MPyV VLP. However, this approach resulted in externalisation of the GFP (Abbing et al 2004;Boura et al 2005;Pleckaityte et al 2015).On the other hand, fusion at the C-terminus of the C-terminal binding fragment of VP2 (VP2C) of SV40 and HPyV allowed encapsidation of GFP inside MPyV VLPs (Inoue et al 2008;Pleckaityte et al 2015).…”

Section: Encapsidation Is Achievable By Utilising a Segment Of The C-mentioning

confidence: 99%

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Murine polyomavirus VLPs as a platform for cytotoxic T cell epitope-based influenza vaccine candidates

Abidin¹

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This research examined the ability of virus-like particles (VLP) assembled from the coat proteins of Murine polyomavirus (MPyV) to carry cytotoxic T cell (Tc) epitopes. MPyV VLPs can be assembled in vitro from purified subunits composed of pentamers of the major coat protein VP1 called capsomeres; and this approach provides fine control over VLP composition and structure. Tc-epitopes are often highly hydrophobic, which poses a significant bioengineering challenge and two distinct approaches were pursued to determine the optimal delivery strategy of Influenza Tc-epitopes by MPyV VLP. Firstly, identified epitopes were externally presented using established sites for peptide insertion on the MPyV major coat protein, VP1. Secondly, polypeptides possessing multiple epitopes were encapsidated within VP1 VLP using precise elements of the minor coat protein VP2/3 that interact with the interior cavity of capsomeres. Hydrophobicity-related capsomere aggregation arising from single Tc epitope presentation was successfully circumvented by engineering charged amino acids (DD) flanking the epitope displayed on a surface-exposed loop of VP1 and by use of an optimised buffer condition. A multiparametric, high-throughput buffer screen was implemented to optimise pH and buffer additives during affinity tag removal. Stable modified capsomere recovered from this reaction were assembly competent in the presence of L-Arginine, and VLP yield was high when modular capsomeres were co-assembled with unmodified VP1 capsomeres forming stable mosaic VLPs. The ability of the MPyV VLP to encapsidate foreign protein was first evaluated and optimised with green fluorescent protein (GFP) as a model protein to enable monitoring and analysis.A minimal VP1-interacting sequence was defined from the carboxy-terminus of VP2/3 (VP2C) and fused to the amino-terminus of GFP, ensuring internalisation of the reporter protein. VP1:VP2C-GFP capsomere complexes were successfully recovered when VP1 was co-expressed with VP2C-GFP, whereas complex formation was not observed when VP1 and VP2C-GFP were mixed in vitro.Recovered capsomere complexes were assembly competent and amount of encapsidated GFP was controllable when VP1:VP2C-GFP capsomere complexes were co-assembled with unmodified VP1 capsomeres in a defined molar ratio. The encapsidation approach was then applied to a subdomain of the Influenza matrix protein M1 by co-expressing VP1 with VP2C-M1-I. M1-I capsomere complexes were assembly competent as indicated by gel electrophoresis, dynamic light scattering, and transmission electron microscopy. To enable recovery of VLPs for analysis and characterisation as well as to confirm encapsidation and the formation of mosaic VLPs, a semi-preparative size exclusion chromatography method was developed. This research was able to address bioengineering challenges posed by hydrophobic epitope presentation and developed MPyV

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Construction of polyomavirus-derived pseudotype virus-like particles displaying a functionally active neutralizing antibody against hepatitis B virus surface antigen

Cited by 22 publications

References 37 publications

Yeast as an expression system for producing virus-like particles: what factors do we need to consider?

Yeast as an expression system for producing virus-like particles: what factors do we need to consider?

Virus-Like Particles as an Instrument of Vaccine Production

Murine polyomavirus VLPs as a platform for cytotoxic T cell epitope-based influenza vaccine candidates

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